DOI: 10.5061/dryad.rr4xgxdfk
title: Data from: Divergent recruitment of developmentally defined neuronal ensembles supports memory dynamics

description: Memories are dynamic constructs whose properties change with time and
 experience. The biological mechanisms underpinning these dynamics remain
 elusive, particularly concerning how shifts in the composition of
 memory-encoding neuronal ensembles influence a memory properties'
 evolution over time. By leveraging a developmental approach to target
 distinct subpopulations of principal neurons, we show that memory encoding
 results in the concurrent establishment of multiple memory traces in the
 mouse hippocampus. Two of these traces are instantiated in subpopulations
 of early- and late-born neurons and follow distinct reactivation
 trajectories post-encoding. Notably, the divergent recruitment of these
 subpopulations underpins memory ensembles’ gradual reorganization, and
 modulates memory persistence and plasticity across multiple learning
 episodes. Thus, our findings reveal profound and intricate relationships
 between ensemble dynamics and memories’ progression over time.
description: # Divergent Recruitment of Developmentally-Defined Neuronal Ensembles
 Supports Memory Dynamics
 [https://doi.org/10.5061/dryad.rr4xgxdfk](https://doi.org/10.5061/dryad.rr4xgxdfk) This dataset contains behavioural, histological, and calcium activity of a study aimed at dissecting the role of Early Born Neurons (EBNs) and Late Born Neurons (LBNs) from hippocampal area CA3 to memory encoding, processing, and recall. ## Description of the data and file structure The data are tabulated in Excel files, each tab refers to a different variable measured and/or a different experiment. Within the table, a brief legend indicates the nature of the data presented. The matlab files report binary traces from calcium imaging experiments. Each file identifier corresponds to an individual animal from the experiment. The file contains two numerical matrices: one with dF/F of every co-recorded neuron alogned throughput the experiments, and one indicating the boundaries (frames numbers) of each individual recordings, which are now concatenated in one singe matrix (frames x neurons). ## Code/Software All files are in .mat and .xls format ## **Content:** Data underlying Figs. 1-5, and Figs. S1-S17. ## **Description of the data and file structure:** Data underlying Figs. 1, 3-5, Figs. S1-S3, Figs. S8-S17 are provided in .xlsx file format. Data underlying Fig 2, and Figs. S4-S7 are provided in .mat file format. File’s structure is as follows:  **adk0997_Figure1.xls**: data underlying Fig. 1 in the main text. This file contains seven tabs.  Figure 1B: Each cell indicates the fraction of CA3 neurons (NeuN+) that are BrdU+ as a function of the day of BrdU Injection. Every cell is a different mouse, Embryonic day of BrdU injection is expressed in the first row. Column A: Injection on E12 Column B: Injection on E13 Column C: Injection on E14 Column D: Injection on E15 Column E: Injection on E16  Figure 1D EBNs: Fraction of c-Fos+ EBNs (BrdU+c-Fos+/BrdU+) upon the Acquisition and Recall of a cFC memory. Every cell is a different mouse, behavioral condition is spelled in row 1. Column A: Home Cage Column B: Acquisition Column C: Recall 1d Column D: Recall 2d Column E: Recall 3d Column F: Recall 4d Column G: Recall 7d Column H: Recall 14d Column I: Recall 28d  Figure 1D LBNs: Fraction of c-Fos+ LBNs (BrdU+c-Fos+/BrdU+) upon the Acquisition and Recall of a cFC memory. Every cell is a different mouse, behavioral condition is spelled in row 1. Column A: Home Cage Column B: Acquisition Column C: Recall 1d Column D: Recall 2d Column E: Recall 3d Column F: Recall 4d Column G: Recall 7d Column H: Recall 14d Column I: Recall 28d  Figure 1E EBNs: Fold difference to chance in the probability of recruitment of EBNs to c-Fos+ ensembles (BrdU+c-Fos+/c-Fos+) upon the Acquisition and Recall of a cFC memory. Every cell is a different mouse, behavioral condition is spelled in row 1. Column A: Home Cage Column B: Acquisition Column C: Recall 1d Column D: Recall 2d Column E: Recall 3d Column F: Recall 4d Column G: Recall 7d Column H: Recall 14d Column I: Recall 28d  Figure 1E LBNs: Fold difference to chance in the probability of recruitment of LBNs to c-Fos+ ensembles (BrdU+c-Fos+/c-Fos+) upon the Acquisition and Recall of a cFC memory. Every cell is a different mouse, behavioral condition is spelled in row 1. Column A: Home Cage Column B: Acquisition Column C: Recall 1d Column D: Recall 2d Column E: Recall 3d Column F: Recall 4d Column G: Recall 7d Column H: Recall 14d Column I: Recall 28d  Figure 1H EB Engram: Fold difference to chance in the probability of triple positive tdTomato+,c-Fos+,BrdU+ Early-Born Engram neurons upon the encoding and retrieval of a cFC memory. Every cell is a different mouse, behavioral condition is spelled in row 1. Column A: Home Cage Column B: Recall 1d Column C: Recall 2d Column D: Recall 3d Column E: Recall 4d Column F: Recall 28d  Figure 1H LB Engram: Fold difference to chance in the probability of triple positive tdTomato+,c-Fos+,BrdU+ Late-Born Engram neurons upon the encoding and retrieval of a cFC memory. Every cell is a different mouse, behavioral condition is spelled in row 1. Column A: Home Cage Column B: Recall 1d Column C: Recall 2d Column D: Recall 3d Column E: Recall 4d Column F: Recall 28d  **adk0997_Figure3.xls**: data underlying Fig. 3 in the main text. This file contains eight tabs.  Figure 3D left: Percentage of time spent freezing in mice where EBNs or LBNs were silenced via CNO injection at cFC Recall 1d. Every cell is a different mouse. Experimental group is indicated in row 1. Column A: Controls Column B: EBNs CNO Column C: LBNs CNO  Figure 3D right: Fraction of CA3 neurons expressing c-Fos (c-Fos+/NeuN+) in mice where EBNs or LBNs were silenced via CNO injection at cFC Recall 1d. Every cell is a different mouse. Experimental group is indicated in row 1. Column A: Controls Column B: EBNs CNO Column C: LBNs CNO  Figure 3E Upper row: Correlation between DREADD expression in EBN-targeted animals (DREADD+/NeuN+) and freezing upon Recall 1d when CNO was delivered 30 minutes before the recall session. Each row is an individual mouse. Experimental group is indicated in row 1. Column A: DREADD+/NeuN+ Column B: Freezing  Figure 3E Lower row: Correlation between DREADD expression in LBN-targeted animals (DREADD+/NeuN+) and freezing upon Recall 1d when CNO was delivered 30 minutes before the recall session. Each row is an individual mouse. Experimental group is indicated in row 1. Column A: DREADD+/NeuN+ Column B: Freezing  Figure 3F left: Percentage of time spent freezing in mice where EBNs or LBNs were silenced via CNO injection at cFC Recall 28d. Every cell is a different mouse. Experimental group is indicated in row 1. Column A: Controls Column B: EBNs CNO Column C: LBNs CNO  Figure 3F right: Fraction of CA3 neurons expressing c-Fos (c-Fos+/NeuN+) in mice where EBNs or LBNs were silenced via CNO injection at cFC Recall 28d. Every cell is a different mouse. Experimental group is indicated in row 1. Column A: Controls Column B: EBNs CNO Column C: LBNs CNO  Figure 3G Upper row: Correlation between DREADD expression in EBN-targeted animals (DREADD+/NeuN+) and freezing upon Recall 28d when CNO was delivered 30 minutes before the recall session. Each row is an individual mouse. Experimental group is indicated in row 1. Column A: DREADD+/NeuN+ Column B: Freezing  Figure 3G Lower row: Correlation between DREADD expression in LBN-targeted animals (DREADD+/NeuN+) and freezing upon Recall 28d when CNO was delivered 30 minutes before the recall session. Each row is an individual mouse. Experimental group is indicated in row 1. Column A: DREADD+/NeuN+ Column B: Freezing **adk0997_Figure4.xls**: data underlying Fig. 4 in the main text. This file contains twelve tabs. Figure 4C EBNs: Fraction of BrdU+ EBNs that are also c-Fos+ in animals where LBNs were expressing DREADDs, with or without the injection of CNO. Every cell is a different mouse. Experimental group is indicated in row 1. Column A: no CNO Column B: CNO Figure 4C LBNs: Fraction of BrdU+ LBNs that are also c-Fos+ in animals where EBNs were expressing DREADDs, with or without the injection of CNO. Every cell is a different mouse. Experimental group is indicated in row 1. Column A: no CNO Column B: CNO  Figure 4E left: Percentage of time spent freezing during the Recall 1d session in mice where EBNs or LBNs were silenced via CNO injection at cFC Acquisition. Every cell is a different mouse. Experimental group is indicated in row 1. Column A: Controls Column B: EBNs CNO Column C: LBNs CNO  Figure 4E right: Fraction of CA3 neurons expressing c-Fos (c-Fos+/NeuN+) upon Recall 1d in mice where EBNs or LBNs were silenced via CNO injection at cFC Acquisition. Every cell is a different mouse. Experimental group is indicated in row 1. Column A: Controls Column B: EBNs CNO Column C: LBNs CNO  Figure 4F left: Percentage of time spent freezing during the Recall 28d session in mice where EBNs or LBNs were silenced via CNO injection at cFC Acquisition. Every cell is a different mouse. Experimental group is indicated in row 1. Column A: Controls Column B: EBNs CNO Column C: LBNs CNO  Figure 4F right: Fraction of CA3 neurons expressing c-Fos (c-Fos+/NeuN+) upon Recall 28d in mice where EBNs or LBNs were silenced via CNO injection at cFC Acquisition. Every cell is a different mouse. Experimental group is indicated in row 1. Column A: Controls Column B: EBNs CNO Column C: LBNs CNO  Figure 4H EBNs: Fraction of c-Fos+ EBNs (BrdU+c-Fos+/BrdU+) upon the Acquisition and during consolidation of a cFC memory. Every cell is a different mouse, behavioral condition is spelled in row 1. Column A: Home Cage Column B: Acquisition Column C: Home 3h Column D: Home 6h Column E: Home 9h Column F: Home 15h Column G: Home 24h  Figure 4H LBNs: Fraction of c-Fos+ LBNs (BrdU+c-Fos+/BrdU+) upon the Acquisition and during consolidation of a cFC memory. Every cell is a different mouse, behavioral condition is spelled in row 1. Column A: Home Cage Column B: Acquisition Column C: Home 3h Column D: Home 6h Column E: Home 9h Column F: Home 15h Column G: Home 24h  Figure 4J left: Percentage of time spent freezing during the Recall 1d session in mice where EBNs or LBNs were silenced via CNO injection during the second window of c-Fos expression during consolidation (15h after Acquisition). Every cell is a different mouse. Experimental group is indicated in row 1. Column A: Controls Column B: EBNs CNO Column C: LBNs CNO  Figure 4J right: Fraction of c-Fos+ neurons in CA3 (c-Fos+/NeuN+) upon the Recall 1d session in mice where EBNs or LBNs were silenced via CNO injection during the second window of c-Fos expression during consolidation (15h after Acquisition). Every cell is a different mouse. Experimental group is indicated in row 1. Column A: Controls Column B: EBNs CNO Column C: LBNs CNO  Figure 4K left: Percentage of time spent freezing during the Recall 28d session in mice where EBNs or LBNs were silenced via CNO injection during the second window of c-Fos expression during consolidation (15h after Acquisition). Every cell is a different mouse. Experimental group is indicated in row 1. Column A: Controls Column B: EBNs CNO Column C: LBNs CNO  Figure 4K right: Fraction of c-Fos+ neurons in CA3 (c-Fos+/NeuN+) upon the Recall 28d session in mice where EBNs or LBNs were silenced via CNO injection during the second window of c-Fos expression during consolidation (15h after Acquisition). Every cell is a different mouse. Experimental group is indicated in row 1. Column A: Controls Column B: EBNs CNO Column C: LBNs CNO  **adk0997_Figure5.xls**: data underlying Fig. 5 in the main text. This file contains four tabs.  Figure 5B: Percentage of time spent freezing in mice trained in the spaced cFC task at different ISI (Inter Session Intervals). Each row pair (A-B; D-E; G-H; J-K) is a repeated longitudinal measure from one individual mouse. Behavioral protocol described in row 1, Training session described in row 2 Column A: % of time spent freezing during Re-Training for ISI=1d Column B: % of time spent freezing during Recall for ISI=1d Column D: % of time spent freezing during Re-Training for ISI=2d Column E: % of time spent freezing during Recall for ISI=2d Column G: % of time spent freezing during Re-Training for ISI=3d Column H: % of time spent freezing during Recall for ISI=3d Column J: % of time spent freezing during Re-Training for ISI=4d Column K: % of time spent freezing during Recall for ISI=4d  Figure 5D: Percentage of time spent freezing in mice trained in the spaced cFC task with ISI=2d, in which EBNs or LBNs were stimulated during Re-Training. Each row pair (A-B; D-E; G-H) is a repeated longitudinal measure from one individual mouse. Experimental group described in row 1, Training session described in row 2 Column A: % of time spent freezing during Re-Training in Controls Column B: % of time spent freezing during Recall in Controls Column D: % of time spent freezing during Re-Training in EBNs CNO Column E: % of time spent freezing during Recall in EBNs CNO Column G: % of time spent freezing during Re-Training in LBNs CNO Column H: % of time spent freezing during Recall in LBNs CNO  Figure 5E: Percentage of time spent freezing in mice trained in the spaced cFC task with ISI=3d, in which EBNs or LBNs were stimulated during Re-Training. Each row pair (A-B; D-E; G-H) is a repeated longitudinal measure from one individual mouse. Experimental group described in row 1, Training session described in row 2 Column A: % of time spent freezing during Re-Training in Controls Column B: % of time spent freezing during Recall in Controls Column D: % of time spent freezing during Re-Training in EBNs CNO Column E: % of time spent freezing during Recall in EBNs CNO Column G: % of time spent freezing during Re-Training in LBNs CNO Column H: % of time spent freezing during Recall in LBNs CNO  Figure 5F: Percentage of time spent freezing in mice trained in the spaced cFC task with ISI=4d, in which EBNs or LBNs were stimulated during Re-Training. Each row pair (A-B; D-E; G-H) is a repeated longitudinal measure from one individual mouse. Experimental group described in row 1, Training session described in row 2 Column A: % of time spent freezing during Re-Training in Controls Column B: % of time spent freezing during Recall in Controls Column D: % of time spent freezing during Re-Training in EBNs CNO Column E: % of time spent freezing during Recall in EBNs CNO Column G: % of time spent freezing during Re-Training in LBNs CNO Column H: % of time spent freezing during Recall in LBNs CNO  **adk0997_FigureS1.xls**: data underlying Fig. S1 in the main text. This file contains twenty-five tabs.  Fig S1A: Percentage of time spent freezing during the Acquisition and Recall of a cFC memory. Every cell is a different mouse, behavioral condition is spelled in row 1. Column A: Naive Column B: Recall 1d Column C: Recall 2d Column D: Recall 3d Column E: Recall 4d Column F: Recall 7d Column G: Recall 14d Column H: Recall 28d  Fig S1C: Fraction of c-Fos+ neurons (c-Fos+/NeuN+) in CA3 upon the Acquisition and Recall of a cFC memory. Every cell is a different mouse, behavioral condition is spelled in row 1. Column A: Home Cage Column B: Acquisition Column C: Recall 1d Column D: Recall 2d Column E: Recall 3d Column F: Recall 4d Column G: Recall 7d Column H: Recall 14d Column I: Recall 28d  Fig S1E: Percentage of time spent freezing during the Acquisition and Recall sessions of a Context Only experiment. Every cell is a different mouse, behavioral condition is spelled in row 1. Column A: Naïve Column B: Recall 1d Column C: Recall 28d  Fig S1F: Fraction of c-Fos+ neurons (c-Fos+/NeuN+) in CA3 upon the Acquisition and Recall of a Context Only memory. Every cell is a different mouse, behavioral condition is spelled in row 1. Column A: Home Cage Column B: Acquisition Column C: Recall 1d Column D: Recall 28d  Fig S1G EBNs: Fraction of c-Fos+ EBNs (BrdU+c-Fos+/BrdU+) upon the Acquisition and Recall of a Context Only memory. Every cell is a different mouse, behavioral condition is spelled in row 1. Column A: Home Cage Column B: Acquisition Column C: Recall 1d Column D: Recall 28d  Fig S1G LBNs: Fraction of c-Fos+ LBNs (BrdU+c-Fos+/BrdU+) upon the Acquisition and Recall of a Context Only memory. Every cell is a different mouse, behavioral condition is spelled in row 1. Column A: Home Cage Column B: Acquisition Column C: Recall 1d Column D: Recall 28d  Fig S1H EBNs: Fold difference to chance in the probability of recruitment of EBNs to c-Fos+ ensembles (BrdU+c-Fos+/c-Fos+) upon the Acquisition and Recall of a Context Only memory. Every cell is a different mouse, behavioral condition is spelled in row 1. Column A: Home Cage Column B: Acquisition Column C: Recall 1d Column D: Recall 28d  Fig S1H LBNs: Fold difference to chance in the probability of recruitment of LBNs to c-Fos+ ensembles (BrdU+c-Fos+/c-Fos+) upon the Acquisition and Recall of a Context Only memory. Every cell is a different mouse, behavioral condition is spelled in row 1. Column A: Home Cage Column B: Acquisition Column C: Recall 1d Column D: Recall 28d  Fig S1J: Fraction of c-Fos+ neurons (c-Fos+/NeuN+) in CA3 upon the Acquisition and Recall of a No Recall memory. Every cell is a different mouse, behavioral condition is spelled in row 1. Column A: Home Cage Column B: Acquisition Column C: Recall 1d Column D: Recall 28d  Fig S1K EBNs Fraction of c-Fos+ EBNs (BrdU+c-Fos+/BrdU+) upon the Acquisition and Recall of a No Recall memory. Every cell is a different mouse, behavioral condition is spelled in row 1. Column A: Home Cage Column B: Acquisition Column C: Recall 1d Column D: Recall 28d  Fig S1K LBNs: Fraction of c-Fos+ LBNs (BrdU+c-Fos+/BrdU+) upon the Acquisition and Recall of a No Recall memory. Every cell is a different mouse, behavioral condition is spelled in row 1. Column A: Home Cage Column B: Acquisition Column C: Recall 1d Column D: Recall 28d  Fig S1L EBNs: Fold difference to chance in the probability of recruitment of EBNs to c-Fos+ ensembles (BrdU+c-Fos+/c-Fos+) upon the Acquisition and Recall of a No Recall memory. Every cell is a different mouse, behavioral condition is spelled in row 1. Column A: Home Cage Column B: Acquisition Column C: Recall 1d Column D: Recall 28d  Fig S1L LBNs: Fold difference to chance in the probability of recruitment of LBNs to c-Fos+ ensembles (BrdU+c-Fos+/c-Fos+) upon the Acquisition and Recall of a No Recall memory. Every cell is a different mouse, behavioral condition is spelled in row 1. Column A: Home Cage Column B: Acquisition Column C: Recall 1d Column D: Recall 28d Fig S1N: Percentage of time spent freezing during the Acquisition and Recall sessions of a Context B experiment. Every cell is a different mouse, behavioral condition is spelled in row 1. Column A: Naïve Column B: Recall 1d Column C: Recall 28d  Fig S1O: Fraction of c-Fos+ neurons (c-Fos+/NeuN+) in CA3 upon the Acquisition and Recall of a Context B memory. Every cell is a different mouse, behavioral condition is spelled in row 1. Column A: Home Cage Column B: Acquisition Column C: Recall 1d Column D: Recall 28d  Fig S1P EBNs: Fraction of c-Fos+ EBNs (BrdU+c-Fos+/BrdU+) upon the Acquisition and Recall of a Context B memory. Every cell is a different mouse, behavioral condition is spelled in row 1. Column A: Home Cage Column B: Acquisition Column C: Recall 1d Column D: Recall 28d  Fig S1P LBNs: Fraction of c-Fos+ LBNs (BrdU+c-Fos+/BrdU+) upon the Acquisition and Recall of a Context B memory. Every cell is a different mouse, behavioral condition is spelled in row 1. Column A: Home Cage Column B: Acquisition Column C: Recall 1d Column D: Recall 28d Fig S1Q EBNs: Fold difference to chance in the probability of recruitment of EBNs to c-Fos+ ensembles (BrdU+c-Fos+/c-Fos+) upon the Acquisition and Recall of a Context B memory. Every cell is a different mouse, behavioral condition is spelled in row 1. Column A: Home Cage Column B: Acquisition Column C: Recall 1d Column D: Recall 28d  Fig S1Q LBNs: Fold difference to chance in the probability of recruitment of LBNs to c-Fos+ ensembles (BrdU+c-Fos+/c-Fos+) upon the Acquisition and Recall of a Context B memory. Every cell is a different mouse, behavioral condition is spelled in row 1. Column A: Home Cage Column B: Acquisition Column C: Recall 1d Column D: Recall 28d  Fig S1S: Percentage of time spent freezing during the Acquisition and Recall of a tFC memory. Every cell is a different mouse, behavioral condition is spelled in row 1. Column A: Naive Column B: Tone Recall 1d Column C: Context Recall 1d Column D: Tone Recall 28d Column E: Context Recall 28d  Fig S1T: Fraction of c-Fos+ neurons (c-Fos+/NeuN+) in CA3 upon the Acquisition and Recall of a tFC memory. Every cell is a different mouse, behavioral condition is spelled in row 1. Column A: Home Cage Column B: Acquisition Column C: Tone Recall 1d Column D: Context Recall 1d Column E: Tone Recall 28d Column F: Context Recall 28d Fig S1U EBNs: Fraction of c-Fos+ EBNs (BrdU+c-Fos+/BrdU+) upon the Acquisition and Recall of a tFC memory. Every cell is a different mouse, behavioral condition is spelled in row 1. Column A: Home Cage Column B: Acquisition Column C: Tone Recall 1d Column D: Context Recall 1d Column E: Tone Recall 28d Column F: Context Recall 28d Fig S1U LBNs: Fraction of c-Fos+ LBNs (BrdU+c-Fos+/BrdU+) upon the Acquisition and Recall of a tFC memory. Every cell is a different mouse, behavioral condition is spelled in row 1. Column A: Home Cage Column B: Acquisition Column C: Tone Recall 1d Column D: Context Recall 1d Column E: Tone Recall 28d Column F: Context Recall 28d  Fig S1V EBNs: Fold difference to chance in the probability of recruitment of EBNs to c-Fos+ ensembles (BrdU+c-Fos+/c-Fos+) upon the Acquisition and Recall of a tFC memory. Every cell is a different mouse, behavioral condition is spelled in row 1. Column A: Home Cage Column B: Acquisition Column C: Tone Recall 1d Column D: Context Recall 1d Column E: Tone Recall 28d Column F: Context Recall 28d Fig S1V LBNs: Fold difference to chance in the probability of recruitment of LBNs to c-Fos+ ensembles (BrdU+c-Fos+/c-Fos+) upon the Acquisition and Recall of a tFC memory. Every cell is a different mouse, behavioral condition is spelled in row 1. Column A: Home Cage Column B: Acquisition Column C: Tone Recall 1d Column D: Context Recall 1d Column E: Tone Recall 28d Column F: Context Recall 28d **adk0997_FigureS2.xls**: data underlying Fig. S2 in the main text. This file contains thirty-one tabs.  Fig S2A: Fraction of NeuN+ neurons that are BrdU+ as a function of the day of BrdU injection. Embryonic day of BrdU injection on the second row, specific area in the first row. Column A: CA3, Injection E12 Column B: CA3, Injection E13 Column C: CA3, Injection E14 Column D: CA3, Injection E15 Column E: CA3, Injection E16 Column G: CA1, Injection E12 Column H: CA1, Injection E13 Column I: CA1, Injection E14 Column J: CA1, Injection E15 Column K: CA1, Injection E16 Column M: Sub, Injection E12 Column N: Sub, Injection E13 Column O: Sub, Injection E14 Column P: Sub, Injection E15 Column Q: Sub, Injection E16 Column S: BLA, Injection E12 Column T: BLA, Injection E13 Column U: BLA, Injection E14 Column V: BLA, Injection E15 Column W: BLA, Injection E16 Column Y: LA, Injection E12 Column Z: LA, Injection E13 Column AA: LA, Injection E14 Column AB: LA, Injection E15 Column AC: LA, Injection E16 Column AE: CeA, Injection E12 Column AF: CeA, Injection E13 Column AG: CeA, Injection E14 Column AH: CeA, Injection E15 Column AI: CeA, Injection E16 Fig S2B CA3 c-Fos: Fraction of c-Fos+ neurons (c-Fos+/NeuN+) in CA3 upon the Acquisition and Recall of a cFC memory. Every cell is a different mouse, behavioral condition is spelled in row 1. Column A: Home Cage Column B: Acquisition Column C: Recall 1d Column D: Recall 28d Fig S2B CA3 EBNs: Fractions of c-Fos+ EBNs (BrdU+c-Fos+/BrdU+, Columns A-D), and fold differences to chance in the probability of recruitment to c-Fos+ ensembles (BrdU+c-Fos+/c-Fos+, Columns F-I) upon the Acquisition and Recall of a cFC memory. Every cell is a different mouse, behavioral condition is spelled in row 2. Column A: Fractions Home Cage Column B: Fractions Acquisition Column C: Fractions Recall 1d Column D: Fractions Recall 28d Column F: Fold differences Home Cage Column G: Fold differences Acquisition Column H: Fold differences Recall 1d Column I: Fold differences Recall 28d Fig S2B CA3 IBNs-13: Fractions of c-Fos+ IBNs-13 (BrdU+c-Fos+/BrdU+, Columns A-D), and fold differences to chance in the probability of recruitment to c-Fos+ ensembles (BrdU+c-Fos+/c-Fos+, Columns F-I) upon the Acquisition and Recall of a cFC memory. Every cell is a different mouse, behavioral condition is spelled in row 2. Column A: Fractions Home Cage Column B: Fractions Acquisition Column C: Fractions Recall 1d Column D: Fractions Recall 28d Column F: Fold differences Home Cage Column G: Fold differences Acquisition Column H: Fold differences Recall 1d Column I: Fold differences Recall 28d Fig S2B CA3 IBNs-14: Fractions of c-Fos+ IBNs-14 (BrdU+c-Fos+/BrdU+, Columns A-D), and fold differences to chance in the probability of recruitment to c-Fos+ ensembles (BrdU+c-Fos+/c-Fos+, Columns F-I) upon the Acquisition and Recall of a cFC memory. Every cell is a different mouse, behavioral condition is spelled in row 2. Column A: Fractions Home Cage Column B: Fractions Acquisition Column C: Fractions Recall 1d Column D: Fractions Recall 28d Column F: Fold differences Home Cage Column G: Fold differences Acquisition Column H: Fold differences Recall 1d Column I: Fold differences Recall 28d Fig S2B CA3 LBNs: Fractions of c-Fos+ LBNs (BrdU+c-Fos+/BrdU+, Columns A-D), and fold differences to chance in the probability of recruitment to c-Fos+ ensembles (BrdU+c-Fos+/c-Fos+, Columns F-I) upon the Acquisition and Recall of a cFC memory. Every cell is a different mouse, behavioral condition is spelled in row 2. Column A: Fractions Home Cage Column B: Fractions Acquisition Column C: Fractions Recall 1d Column D: Fractions Recall 28d Column F: Fold differences Home Cage Column G: Fold differences Acquisition Column H: Fold differences Recall 1d Column I: Fold differences Recall 28d Fig S2C CA1 c-Fos: Fraction of c-Fos+ neurons (c-Fos+/NeuN+) in CA1 upon the Acquisition and Recall of a cFC memory. Every cell is a different mouse, behavioral condition is spelled in row 1. Column A: Home Cage Column B: Acquisition Column C: Recall 1d Column D: Recall 28d Fig S2C CA1 EBNs: Fractions of c-Fos+ EBNs (BrdU+c-Fos+/BrdU+, Columns A-D), and fold differences to chance in the probability of recruitment to c-Fos+ ensembles (BrdU+c-Fos+/c-Fos+, Columns F-I) upon the Acquisition and Recall of a cFC memory. Every cell is a different mouse, behavioral condition is spelled in row 2. Column A: Fractions Home Cage Column B: Fractions Acquisition Column C: Fractions Recall 1d Column D: Fractions Recall 28d Column F: Fold differences Home Cage Column G: Fold differences Acquisition Column H: Fold differences Recall 1d Column I: Fold differences Recall 28d Fig S2C CA1 IBNs-13: Fractions of c-Fos+ IBNs-13 (BrdU+c-Fos+/BrdU+, Columns A-D), and fold differences to chance in the probability of recruitment to c-Fos+ ensembles (BrdU+c-Fos+/c-Fos+, Columns F-I) upon the Acquisition and Recall of a cFC memory. Every cell is a different mouse, behavioral condition is spelled in row 2. Column A: Fractions Home Cage Column B: Fractions Acquisition Column C: Fractions Recall 1d Column D: Fractions Recall 28d Column F: Fold differences Home Cage Column G: Fold differences Acquisition Column H: Fold differences Recall 1d Column I: Fold differences Recall 28d Fig S2C CA1 IBNs-14: Fractions of c-Fos+ IBNs-14 (BrdU+c-Fos+/BrdU+, Columns A-D), and fold differences to chance in the probability of recruitment to c-Fos+ ensembles (BrdU+c-Fos+/c-Fos+, Columns F-I) upon the Acquisition and Recall of a cFC memory. Every cell is a different mouse, behavioral condition is spelled in row 2. Column A: Fractions Home Cage Column B: Fractions Acquisition Column C: Fractions Recall 1d Column D: Fractions Recall 28d Column F: Fold differences Home Cage Column G: Fold differences Acquisition Column H: Fold differences Recall 1d Column I: Fold differences Recall 28d Fig S2C CA1 LBNs: Fractions of c-Fos+ LBNs (BrdU+c-Fos+/BrdU+, Columns A-D), and fold differences to chance in the probability of recruitment to c-Fos+ ensembles (BrdU+c-Fos+/c-Fos+, Columns F-I) upon the Acquisition and Recall of a cFC memory. Every cell is a different mouse, behavioral condition is spelled in row 2. Column A: Fractions Home Cage Column B: Fractions Acquisition Column C: Fractions Recall 1d Column D: Fractions Recall 28d Column F: Fold differences Home Cage Column G: Fold differences Acquisition Column H: Fold differences Recall 1d Column I: Fold differences Recall 28d Fig S2D Sub c-Fos: Fraction of c-Fos+ neurons (c-Fos+/NeuN+) in Sub upon the Acquisition and Recall of a cFC memory. Every cell is a different mouse, behavioral condition is spelled in row 1. Column A: Home Cage Column B: Acquisition Column C: Recall 1d Column D: Recall 28d Fig S2D Sub EBNs: Fractions of c-Fos+ EBNs (BrdU+c-Fos+/BrdU+, Columns A-D), and fold differences to chance in the probability of recruitment to c-Fos+ ensembles (BrdU+c-Fos+/c-Fos+, Columns F-I) upon the Acquisition and Recall of a cFC memory. Every cell is a different mouse, behavioral condition is spelled in row 2. Column A: Fractions Home Cage Column B: Fractions Acquisition Column C: Fractions Recall 1d Column D: Fractions Recall 28d Column F: Fold differences Home Cage Column G: Fold differences Acquisition Column H: Fold differences Recall 1d Column I: Fold differences Recall 28d Fig S2D Sub IBNs-13: Fractions of c-Fos+ IBNs-13 (BrdU+c-Fos+/BrdU+, Columns A-D), and fold differences to chance in the probability of recruitment to c-Fos+ ensembles (BrdU+c-Fos+/c-Fos+, Columns F-I) upon the Acquisition and Recall of a cFC memory. Every cell is a different mouse, behavioral condition is spelled in row 2. Column A: Fractions Home Cage Column B: Fractions Acquisition Column C: Fractions Recall 1d Column D: Fractions Recall 28d Column F: Fold differences Home Cage Column G: Fold differences Acquisition Column H: Fold differences Recall 1d Column I: Fold differences Recall 28d Fig S2D Sub IBNs-14: Fractions of c-Fos+ IBNs-14 (BrdU+c-Fos+/BrdU+, Columns A-D), and fold differences to chance in the probability of recruitment to c-Fos+ ensembles (BrdU+c-Fos+/c-Fos+, Columns F-I) upon the Acquisition and Recall of a cFC memory. Every cell is a different mouse, behavioral condition is spelled in row 2. Column A: Fractions Home Cage Column B: Fractions Acquisition Column C: Fractions Recall 1d Column D: Fractions Recall 28d Column F: Fold differences Home Cage Column G: Fold differences Acquisition Column H: Fold differences Recall 1d Column I: Fold differences Recall 28d Fig S2D Sub LBNs: Fractions of c-Fos+ LBNs (BrdU+c-Fos+/BrdU+, Columns A-D), and fold differences to chance in the probability of recruitment to c-Fos+ ensembles (BrdU+c-Fos+/c-Fos+, Columns F-I) upon the Acquisition and Recall of a cFC memory. Every cell is a different mouse, behavioral condition is spelled in row 2. Column A: Fractions Home Cage Column B: Fractions Acquisition Column C: Fractions Recall 1d Column D: Fractions Recall 28d Column F: Fold differences Home Cage Column G: Fold differences Acquisition Column H: Fold differences Recall 1d Column I: Fold differences Recall 28d Fig S2E BLA c-Fos: Fraction of c-Fos+ neurons (c-Fos+/NeuN+) in BLA upon the Acquisition and Recall of a cFC memory. Every cell is a different mouse, behavioral condition is spelled in row 1. Column A: Home Cage Column B: Acquisition Column C: Recall 1d Column D: Recall 28d Fig S2E BLA EBNs: Fractions of c-Fos+ EBNs (BrdU+c-Fos+/BrdU+, Columns A-D), and fold differences to chance in the probability of recruitment to c-Fos+ ensembles (BrdU+c-Fos+/c-Fos+, Columns F-I) upon the Acquisition and Recall of a cFC memory. Every cell is a different mouse, behavioral condition is spelled in row 2. Column A: Fractions Home Cage Column B: Fractions Acquisition Column C: Fractions Recall 1d Column D: Fractions Recall 28d Column F: Fold differences Home Cage Column G: Fold differences Acquisition Column H: Fold differences Recall 1d Column I: Fold differences Recall 28d Fig S2E BLA IBNs-13: Fractions of c-Fos+ IBNs-13 (BrdU+c-Fos+/BrdU+, Columns A-D), and fold differences to chance in the probability of recruitment to c-Fos+ ensembles (BrdU+c-Fos+/c-Fos+, Columns F-I) upon the Acquisition and Recall of a cFC memory. Every cell is a different mouse, behavioral condition is spelled in row 2. Column A: Fractions Home Cage Column B: Fractions Acquisition Column C: Fractions Recall 1d Column D: Fractions Recall 28d Column F: Fold differences Home Cage Column G: Fold differences Acquisition Column H: Fold differences Recall 1d Column I: Fold differences Recall 28d Fig S2E BLA IBNs-14: Fractions of c-Fos+ IBNs-14 (BrdU+c-Fos+/BrdU+, Columns A-D), and fold differences to chance in the probability of recruitment to c-Fos+ ensembles (BrdU+c-Fos+/c-Fos+, Columns F-I) upon the Acquisition and Recall of a cFC memory. Every cell is a different mouse, behavioral condition is spelled in row 2. Column A: Fractions Home Cage Column B: Fractions Acquisition Column C: Fractions Recall 1d Column D: Fractions Recall 28d Column F: Fold differences Home Cage Column G: Fold differences Acquisition Column H: Fold differences Recall 1d Column I: Fold differences Recall 28d Fig S2E BLA LBNs: Fractions of c-Fos+ LBNs (BrdU+c-Fos+/BrdU+, Columns A-D), and fold differences to chance in the probability of recruitment to c-Fos+ ensembles (BrdU+c-Fos+/c-Fos+, Columns F-I) upon the Acquisition and Recall of a cFC memory. Every cell is a different mouse, behavioral condition is spelled in row 2. Column A: Fractions Home Cage Column B: Fractions Acquisition Column C: Fractions Recall 1d Column D: Fractions Recall 28d Column F: Fold differences Home Cage Column G: Fold differences Acquisition Column H: Fold differences Recall 1d Column I: Fold differences Recall 28d Fig S2F LA c-Fos: Fraction of c-Fos+ neurons (c-Fos+/NeuN+) in LA upon the Acquisition and Recall of a cFC memory. Every cell is a different mouse, behavioral condition is spelled in row 1. Column A: Home Cage Column B: Acquisition Column C: Recall 1d Column D: Recall 28d Fig S2F LA EBNs: Fractions of c-Fos+ EBNs (BrdU+c-Fos+/BrdU+, Columns A-D), and fold differences to chance in the probability of recruitment to c-Fos+ ensembles (BrdU+c-Fos+/c-Fos+, Columns F-I) upon the Acquisition and Recall of a cFC memory. Every cell is a different mouse, behavioral condition is spelled in row 2. Column A: Fractions Home Cage Column B: Fractions Acquisition Column C: Fractions Recall 1d Column D: Fractions Recall 28d Column F: Fold differences Home Cage Column G: Fold differences Acquisition Column H: Fold differences Recall 1d Column I: Fold differences Recall 28d Fig S2F LA IBNs-13: Fractions of c-Fos+ IBNs-13 (BrdU+c-Fos+/BrdU+, Columns A-D), and fold differences to chance in the probability of recruitment to c-Fos+ ensembles (BrdU+c-Fos+/c-Fos+, Columns F-I) upon the Acquisition and Recall of a cFC memory. Every cell is a different mouse, behavioral condition is spelled in row 2. Column A: Fractions Home Cage Column B: Fractions Acquisition Column C: Fractions Recall 1d Column D: Fractions Recall 28d Column F: Fold differences Home Cage Column G: Fold differences Acquisition Column H: Fold differences Recall 1d Column I: Fold differences Recall 28d Fig S2F LA IBNs-14: Fractions of c-Fos+ IBNs-14 (BrdU+c-Fos+/BrdU+, Columns A-D), and fold differences to chance in the probability of recruitment to c-Fos+ ensembles (BrdU+c-Fos+/c-Fos+, Columns F-I) upon the Acquisition and Recall of a cFC memory. Every cell is a different mouse, behavioral condition is spelled in row 2. Column A: Fractions Home Cage Column B: Fractions Acquisition Column C: Fractions Recall 1d Column D: Fractions Recall 28d Column F: Fold differences Home Cage Column G: Fold differences Acquisition Column H: Fold differences Recall 1d Column I: Fold differences Recall 28d Fig S2F LA LBNs: Fractions of c-Fos+ LBNs (BrdU+c-Fos+/BrdU+, Columns A-D), and fold differences to chance in the probability of recruitment to c-Fos+ ensembles (BrdU+c-Fos+/c-Fos+, Columns F-I) upon the Acquisition and Recall of a cFC memory. Every cell is a different mouse, behavioral condition is spelled in row 2. Column A: Fractions Home Cage Column B: Fractions Acquisition Column C: Fractions Recall 1d Column D: Fractions Recall 28d Column F: Fold differences Home Cage Column G: Fold differences Acquisition Column H: Fold differences Recall 1d Column I: Fold differences Recall 28d Fig S2G CeA c-Fos: Fraction of c-Fos+ neurons (c-Fos+/NeuN+) in CeA upon the Acquisition and Recall of a cFC memory. Every cell is a different mouse, behavioral condition is spelled in row 1. Column A: Home Cage Column B: Acquisition Column C: Recall 1d Column D: Recall 28d Fig S2G CeA EBNs: Fractions of c-Fos+ EBNs (BrdU+c-Fos+/BrdU+, Columns A-D), and fold differences to chance in the probability of recruitment to c-Fos+ ensembles (BrdU+c-Fos+/c-Fos+, Columns F-I) upon the Acquisition and Recall of a cFC memory. Every cell is a different mouse, behavioral condition is spelled in row 2. Column A: Fractions Home Cage Column B: Fractions Acquisition Column C: Fractions Recall 1d Column D: Fractions Recall 28d Column F: Fold differences Home Cage Column G: Fold differences Acquisition Column H: Fold differences Recall 1d Column I: Fold differences Recall 28d Fig S2G CeA IBNs-13: Fractions of c-Fos+ IBNs-13 (BrdU+c-Fos+/BrdU+, Columns A-D), and fold differences to chance in the probability of recruitment to c-Fos+ ensembles (BrdU+c-Fos+/c-Fos+, Columns F-I) upon the Acquisition and Recall of a cFC memory. Every cell is a different mouse, behavioral condition is spelled in row 2. Column A: Fractions Home Cage Column B: Fractions Acquisition Column C: Fractions Recall 1d Column D: Fractions Recall 28d Column F: Fold differences Home Cage Column G: Fold differences Acquisition Column H: Fold differences Recall 1d Column I: Fold differences Recall 28d Fig S2G CeA IBNs-14: Fractions of c-Fos+ IBNs-14 (BrdU+c-Fos+/BrdU+, Columns A-D), and fold differences to chance in the probability of recruitment to c-Fos+ ensembles (BrdU+c-Fos+/c-Fos+, Columns F-I) upon the Acquisition and Recall of a cFC memory. Every cell is a different mouse, behavioral condition is spelled in row 2. Column A: Fractions Home Cage Column B: Fractions Acquisition Column C: Fractions Recall 1d Column D: Fractions Recall 28d Column F: Fold differences Home Cage Column G: Fold differences Acquisition Column H: Fold differences Recall 1d Column I: Fold differences Recall 28d Fig S2G CeA LBNs: Fractions of c-Fos+ LBNs (BrdU+c-Fos+/BrdU+, Columns A-D), and fold differences to chance in the probability of recruitment to c-Fos+ ensembles (BrdU+c-Fos+/c-Fos+, Columns F-I) upon the Acquisition and Recall of a cFC memory. Every cell is a different mouse, behavioral condition is spelled in row 2. Column A: Fractions Home Cage Column B: Fractions Acquisition Column C: Fractions Recall 1d Column D: Fractions Recall 28d Column F: Fold differences Home Cage Column G: Fold differences Acquisition Column H: Fold differences Recall 1d Column I: Fold differences Recall 28d  **adk0997_FigureS3.xls**: data underlying Fig. S3 in the main text. This file contains one tab.  Figure S3C: Columns A-F: fold difference to chance in the probability of double positive tdTomato+, NeuN+ neurons; Columns H-K: fold difference to chance in the probability of triple positive tdTomato+,c-Fos+,BrdU+ Early- or Late-Born Engram neurons. Every cell is a different mouse. Behavioral condition of tagging and animal perfusion is indicated in row 1. Column A: 4OHT in Home Cage, perfusion on day 1. Column B: 4OHT in Home Cage, perfusion on day 4. Column C: Saline at Acquisition, perfusion on day 1. Column D: Saline at Acquisition, perfusion on day 4. Column E: 4OHT at Acquisition, perfusion on day 1. Column F: 4OHT at Acquisition, perfusion on day 4. Column H: EB-Engram neurons, 4OHT at Acquisition, perfusion on day 1. Column I: EB-Engram neurons, 4OHT at Acquisition, perfusion on day 4. Column J: LB-Engram neurons, 4OHT at Acquisition, perfusion on day 1. Column K: LB-Engram neurons, 4OHT at Acquisition, perfusion on day 4. **adk0997_FigureS8.xls**: data underlying Fig. S8 in the main text. This file contains eight tabs. Fig S8B: Fraction of CA3 Network (NeuN+) labelled either by BrdU or by DREADDs when these were targeted to EBNs or LBNs. Each cell is a mouse, labelling method and isochronic population targeted are described in row 1. Column A: EBNs BrdU Column B: EBNs DREADD Column C: LBNs BrdU Column D: LBNs DREADD Fig S8D: Fraction of CA3 Network (NeuN+) labelled both by BrdU and DREADDs when these were targeted to EBNs or LBNs in the same animals. Each cell is a mouse, labelling method and isochronic population targeted are described in row 1. Column A: (LBNs)BrdU/(EBNs)mCherry Column B: (LBNs)mCherry/(EBNs)BrdU Fig S8F: Fraction of CA3 Network (NeuN+) expressing c-Fos when activating EBNs or LBNs in the Home Cage. Each cell is a mouse, isochronic population labelled and CNO injection are described in row 1. Column A: Controls Column B: Controls CNO Column C: EBNs Column D: EBNs CNO Column E: LBNs Column F: LBNs CNO Fig S8G: Fraction of DREADD+ neurons expressing c-Fos when DREADDs were targeted to EBNs or LBNs in the Home Cage. Each cell is a mouse, isochronic population labelled and CNO injection are described in row 1. Column A: EBNs Column B: EBNs CNO Column C: LBNs Column D: LBNs CNO Fig S8I: Fraction of CA3 Network (NeuN+) expressing c-Fos upon cFC Acquisition when silencing EBNs or LBNs. Each cell is a mouse, behavioral condition and isochronic population labelled are described in row 1. Column A: Home Cage Column B: Controls Column C: Silencing EBNs Column D: Silencing LBNs Fig S8J: Fraction of DREADDs+ neurons that are also c-Fos+ upon cFC Acquisition with or without the injection of CNO. Every cell is a different mouse. Experimental group is indicated in row 2, Isochronic subpopulation targeted with DREADD in row 1. Column A: DREADD in EBNs, Home Cage Column B: DREADD in EBNs, Acquisition – no CNO Column C: DREADD in EBNs, Acquisition – CNO Column E: DREADD in LBNs, Home Cage Column F: DREADD in LBNs, Acquisition – no CNO Column G: DREADD in LBNs, Acquisition – CNO Fig S8L: Fraction of DREADDs+ neurons that are also c-Fos+ upon cFC Recall 1d with or without the injection of CNO. Every cell is a different mouse. Experimental group is indicated in row 2, Isochronic subpopulation targeted with DREADD in row 1. Column A: DREADD in EBNs, Saline Column B: DREADD in EBNs, Activation Column C: DREADD in EBNs, Inhibition Column E: DREADD in LBNs, Saline Column F: DREADD in LBNs, Activation Column G: DREADD in LBNs, Inhibition Fig S8M: Fraction of DREADDs+ neurons that are also c-Fos+ upon cFC Recall 28d with or without the injection of CNO. Every cell is a different mouse. Experimental group is indicated in row 2, Isochronic subpopulation targeted with DREADD in row 1. Column A: DREADD in EBNs, Saline Column B: DREADD in EBNs, Activation Column C: DREADD in EBNs, Inhibition Column E: DREADD in LBNs, Saline Column F: DREADD in LBNs, Activation Column G: DREADD in LBNs, Inhibition **adk0997_FigureS9.xls**: data underlying Fig. S9 in the main text. This file contains four tabs. Fig S9A Top Row: Correlation between Freezing at Recall 1d and the fraction of mCherry+ neurons in each area (normalized by NeuN+) when DREADD are targeted to EBNs. Each cell is a mouse, Columns A indicates % of time spent freezing at Recall, columns B-E indicate DREADD+ % of neurons in CA1 (B), DG(C), Sub(D), Cortex(E).  Fig S9A Bottom Row: Correlation between Freezing at Recall 1d and the fraction of mCherry+ neurons in each area (normalized by NeuN+) when DREADD are targeted to LBNs. Each cell is a mouse, Columns A indicates % of time spent freezing at Recall, columns B-E indicate DREADD+ % of neurons in CA1 (B), DG(C), Sub(D), Cortex(E).  Fig S9B Top Row: Correlation between Freezing at Recall 28d and the fraction of mCherry+ neurons in each area (normalized by NeuN+) when DREADD are targeted to EBNs. Each cell is a mouse, Columns A indicates % of time spent freezing at Recall, columns B-E indicate DREADD+ % of neurons in CA1 (B), DG(C), Sub(D), Cortex(E).  Fig S9B Bottom Row: Correlation between Freezing at Recall 28d and the fraction of mCherry+ neurons in each area (normalized by NeuN+) when DREADD are targeted to LBNs. Each cell is a mouse, Columns A indicates % of time spent freezing at Recall, columns B-E indicate DREADD+ % of neurons in CA1 (B), DG(C), Sub(D), Cortex(E).  **adk0997_FigureS10.xls**: data underlying Fig. S10 in the main text. This file contains nine tabs.  Fig S10C: Fraction of CA1 DREADDs+ neurons that are also c-Fos+ upon cFC Recall 1d with or without the injection of CNO. Every cell is a different mouse. Experimental group is indicated in row 1. Column A: DREADD in EBNs, Saline Column B: DREADD in EBNs, CNO Column D: DREADD in LBNs, Saline Column E: DREADD in LBNs, CNO  Fig S10D left: Percentage of time spent freezing during the Recall 1d of a cFC memory when EBNs or LBNs are silenced. Every cell is a different mouse, experimental group indicated in row1. Column A: Controls Column B: EBNs CNO Column C: LBNs CNO  Fig S10D right: Fraction of CA1 DREADDs+ neurons that are also c-Fos+ upon cFC Recall 1d when EBNs or LBNs are silenced. Every cell is a different mouse. Experimental group is indicated in row 1. Column A: Controls Column B: EBNs CNO Column C: LBNs CNO  Fig S10E: Fraction of CA1 DREADDs+ neurons that are also c-Fos+ upon cFC Recall 28d with or without the injection of CNO. Every cell is a different mouse. Experimental group is indicated in row 1. Column A: DREADD in EBNs, Saline Column B: DREADD in EBNs, CNO Column D: DREADD in LBNs, Saline Column E: DREADD in LBNs, CNO  Fig S10F left: Percentage of time spent freezing during the Recall 28d of a cFC memory when EBNs or LBNs are silenced. Every cell is a different mouse, experimental group indicated in row1. Column A: Controls Column B: EBNs CNO Column C: LBNs CNO  Fig S10F right: Fraction of CA1 DREADDs+ neurons that are also c-Fos+ upon cFC Recall 28d when EBNs or LBNs are silenced. Every cell is a different mouse. Experimental group is indicated in row 1. Column A: Controls Column B: EBNs CNO Column C: LBNs CNO  Fig S10I: Fraction of dSub DREADDs+ neurons that are also c-Fos+ upon cFC Recall 1d with or without the injection of CNO. Every cell is a different mouse. Experimental group is indicated in row 1. Column A: DREADD in EBNs, Saline Column B: DREADD in EBNs, CNO Column D: DREADD in LBNs, Saline Column E: DREADD in LBNs, CNO Fig S10J left: Percentage of time spent freezing during the Recall 1d of a cFC memory when EBNs or LBNs are silenced. Every cell is a different mouse, experimental group indicated in row1. Column A: Controls Column B: EBNs CNO Column C: LBNs CNO  Fig S10J right: Fraction of CA1 DREADDs+ neurons that are also c-Fos+ upon cFC Recall 1d when EBNs or LBNs are silenced. Every cell is a different mouse. Experimental group is indicated in row 1. Column A: Controls Column B: EBNs CNO Column C: LBNs CNO  **adk0997_FigureS11.xls**: data underlying Fig. S11 in the main text. This file contains six tabs.  Fig S11C: Fraction of Ca3 Arch+ neurons that are also c-Fos+ upon cFC Recall 1d with or without delivery of light. Every cell is a different mouse. Experimental group is indicated in row 1. Column A: Arch in EBNs, Controls Column B: Arch in EBNs, Optp Column D: Arch in LBNs, Controls Column E: Arch in LBNs, Opto  Fig S11D left: Percentage of time spent freezing during the Recall 1d of a cFC memory when EBNs or LBNs are silenced via optogenetics. Every cell is a different mouse, experimental group indicated in row 1. Column A: Controls Column B: EBNs Column C: LBNs  Fig S11D right: Fraction of CA3 neurons (NeuN+) that are also c-Fos+ upon cFC Recall 1d when EBNs or LBNs are silenced with Optogenetics. Every cell is a different mouse. Experimental group is indicated in row 1. Column A: Controls Column B: EBNs Column C: LBNs  Fig S11E: Fraction of Ca3 Arch+ neurons that are also c-Fos+ upon cFC Recall 28d with or without delivery of light. Every cell is a different mouse. Experimental group is indicated in row 1. Column A: Arch in EBNs, Controls Column B: Arch in EBNs, Optp Column D: Arch in LBNs, Controls Column E: Arch in LBNs, Opto  Fig S11F left: Percentage of time spent freezing during the Recall 28d of a cFC memory when EBNs or LBNs are silenced via optogenetics. Every cell is a different mouse, experimental group indicated in row 1. Column A: Controls Column B: EBNs Column C: LBNs  Fig S11F right: Fraction of CA3 neurons (NeuN+) that are also c-Fos+ upon cFC Recall 28d when EBNs or LBNs are silenced with Optogenetics. Every cell is a different mouse. Experimental group is indicated in row 1. Column A: Controls Column B: EBNs Column C: LBNs **adk0997_FigureS12.xls**: data underlying Fig. S12 in the main text. This file contains eight tabs. Fig S12B left: Percentage of time spent freezing during the Recall 1d of a cFC memory when EBNs or LBNs are activated with chemogenetics. Every cell is a different mouse, experimental group indicated in row 1. Column A: Controls Column B: EBNs CNO Column C: LBNs CNO Fig S12B right: Fraction of CA3 neurons (NeuN+) that are also c-Fos+ upon cFC Recall 1d when EBNs or LBNs are activated with chemogenetics. Every cell is a different mouse. Experimental group is indicated in row 1. Column A: Controls Column B: EBNs CNO Column C: LBNs CNO Fig S12C left: Percentage of time spent freezing during the Recall 28d of a cFC memory when EBNs or LBNs are activated with chemogenetics. Every cell is a different mouse, experimental group indicated in row 1. Column A: Controls Column B: EBNs CNO Column C: LBNs CNO Fig S12C right: Fraction of CA3 neurons (NeuN+) that are also c-Fos+ upon cFC Recall 28d when EBNs or LBNs are activated with chemogenetics. Every cell is a different mouse. Experimental group is indicated in row 1. Column A: Controls Column B: EBNs CNO Column C: LBNs CNO Fig S12D: Fraction of CA3 c-Fos+ neurons that are also DREADD+ or DREADD- upon cFC Recall 1d when EBNs or LBNs are activated with chemogenetics. Every cell is a different mouse. Experimental group is indicated in row 2, Isochronic population targeted with DREADD in row 1. Column A: EBNs Saline, Dreadd+c-Fos+/c-Fos+ Column B: EBNs Saline, Dreadd-c-Fos+/c-Fos+ Column D: EBNs CNO, Dreadd+c-Fos+/c-Fos+ Column E: EBNs CNO, Dreadd-c-Fos+/c-Fos+ Column H: LBNs Saline, Dreadd+c-Fos+/c-Fos+ Column I: LBNs Saline, Dreadd-c-Fos+/c-Fos+ Column K: LBNs CNO, Dreadd+c-Fos+/c-Fos+ Column L: LBNs CNO, Dreadd-c-Fos+/c-Fos+ Fig S12E: Fraction of CA3 c-Fos+ neurons that are also DREADD+ or DREADD- upon cFC Recall 28d when EBNs or LBNs are activated with chemogenetics. Every cell is a different mouse. Experimental group is indicated in row 2, Isochronic population targeted with DREADD in row 1. Column A: EBNs Saline, Dreadd+c-Fos+/c-Fos+ Column B: EBNs Saline, Dreadd-c-Fos+/c-Fos+ Column D: EBNs CNO, Dreadd+c-Fos+/c-Fos+ Column E: EBNs CNO, Dreadd-c-Fos+/c-Fos+ Column H: LBNs Saline, Dreadd+c-Fos+/c-Fos+ Column I: LBNs Saline, Dreadd-c-Fos+/c-Fos+ Column K: LBNs CNO, Dreadd+c-Fos+/c-Fos+ Column L: LBNs CNO, Dreadd-c-Fos+/c-Fos+ Fig S12F left: Percentage of time spent freezing during the Recall 2d of a cFC memory when EBNs or LBNs are activated with chemogenetics. Every cell is a different mouse, experimental group indicated in row 1. Column A: Controls Column B: EBNs CNO Column C: LBNs CNO Fig S12F right: Fraction of CA3 neurons (NeuN+) that are also c-Fos+ upon cFC Recall 2d when EBNs or LBNs are activated with chemogenetics. Every cell is a different mouse. Experimental group is indicated in row 1. Column A: Controls Column B: EBNs CNO Column C: LBNs CNO **adk0997_FigureS13.xls**: data underlying Fig. S13 in the main text. This file contains one tab.  Fig S13B: Fraction of time spent freezing in animals trained in the cFC tasks with double injection of CNO. Each cell is one mouse, experimental conditions in row 2, experimental groups in row 1. Column A: Controls, Pre-Shock Column B: Controls, Recall 1d Column C: Controls, Recall 28d Column E: EBNs, Pre-Shock Column F: EBNs, Recall 1d Column G: EBNs, Recall 28d Column E: LBNs, Pre-Shock Column F: LBNs, Recall 1d Column G: LBNs, Recall 28d  **adk0997_FigureS14.xls**: data underlying Fig. S14 in the main text. This file contains four tabs.  Fig S14A left: Correlation between Freezing at Recall 1d and the fraction of mCherry+ neurons in CA3 (normalized by NeuN+) when DREADD are targeted to EBNs and CNO is delivered at Acquisition. Each cell is a mouse, Columns A indicates % of time spent freezing at Recall, columns B indicates % of DREADD+ neurons in CA3.  Fig S14A right: Correlation between Freezing at Recall 1d and the fraction of mCherry+ neurons in CA3 (normalized by NeuN+) when DREADD are targeted to LBNs and CNO is delivered at Acquisition. Each cell is a mouse, Columns A indicates % of time spent freezing at Recall, columns B indicates % of DREADD+ neurons in CA3.  Fig S14B left: Correlation between Freezing at Recall 28d and the fraction of mCherry+ neurons in CA3 (normalized by NeuN+) when DREADD are targeted to EBNs and CNO is delivered at Acquisition. Each cell is a mouse, Columns A indicates % of time spent freezing at Recall, columns B indicates % of DREADD+ neurons in CA3.  Fig S14B right: Correlation between Freezing at Recall 28d and the fraction of mCherry+ neurons in CA3 (normalized by NeuN+) when DREADD are targeted to LBNs and CNO is delivered at Acquisition. Each cell is a mouse, Columns A indicates % of time spent freezing at Recall, columns B indicates % of DREADD+ neurons in CA3.  **adk0997_FigureS15.xls**: data underlying Fig. S15 in the main text. This file contains nine tabs.  Fig S15A: Fraction of c-Fos+ CA3 neurons (c-Fos+/NeuN+) upon the Acquisition and during consolidation of a cFC memory. Every cell is a different mouse, behavioral condition is spelled in row 1. Column A: Home Cage Column B: Acquisition Column C: 3h Column D: 6h Column E: 9h Column F: 15h Column G: 24h  Fig S15B left: Correlation between Freezing at Recall 1d and the fraction of mCherry+ neurons in CA3 (normalized by NeuN+) when DREADD are targeted to EBNs and CNO is delivered at 13.5h after Acquisition. Each row is a mouse, Columns A indicates % of time spent freezing at Recall, columns B indicates % of DREADD+ neurons in CA3.  Fig S15B right: Correlation between Freezing at Recall 1d and the fraction of mCherry+ neurons in CA3 (normalized by NeuN+) when DREADD are targeted to LBNs and CNO is delivered at 13.5h after Acquisition. Each row is a mouse, Columns A indicates % of time spent freezing at Recall, columns B indicates % of DREADD+ neurons in CA3.  Fig S15C left: Correlation between Freezing at Recall 28d and the fraction of mCherry+ neurons in CA3 (normalized by NeuN+) when DREADD are targeted to EBNs and CNO is delivered at 13.5h after Acquisition. Each row is a mouse, Columns A indicates % of time spent freezing at Recall, columns B indicates % of DREADD+ neurons in CA3.  Fig S15C right: Correlation between Freezing at Recall 28d and the fraction of mCherry+ neurons in CA3 (normalized by NeuN+) when DREADD are targeted to LBNs and CNO is delivered at 13.5h after Acquisition. Each row is a mouse, Columns A indicates % of time spent freezing at Recall, columns B indicates % of DREADD+ neurons in CA3.  Fig 15D left: Percentage of time spent freezing in mice where EBNs or LBNs were silenced via CNO injection 7.5 hours after Acquisition (i.e. silencing at 9h during consolidation). Every cell is a different mouse, experimental group is spelled in row 1. Column A: Controls Column B: EBNs CNO Column C: LBNs CNO  Fig 15D right: Fraction of CA3 neurons expressing c-Fos (c-Fos+/NeuN+) in mice where EBNs or LBNs were silenced via CNO injection 7.5 hours after Acquisition (i.e. silencing at 9h during consolidation). Every cell is a different mouse, experimental group is indicated in row 1. Column A: Controls Column B: EBNs CNO Column C: LBNs CNO  Fig S15E left: Correlation between Freezing at Recall 1d and the fraction of mCherry+ neurons in CA3 (normalized by NeuN+) when DREADD are targeted to EBNs and CNO is delivered at at 7.5h after Acquisition. Each row is a mouse, Columns A indicates % of time spent freezing at Recall, columns B indicates % of DREADD+ neurons in CA3.  Fig S15E right: Correlation between Freezing at Recall 1d and the fraction of mCherry+ neurons in CA3 (normalized by NeuN+) when DREADD are targeted to LBNs and CNO is delivered at at 7.5h after Acquisition. Each row is a mouse, Columns A indicates % of time spent freezing at Recall, columns B indicates % of DREADD+ neurons in CA3.  **adk0997_FigureS16.xls**: data underlying Fig. S16 in the main text. This file contains four tabs.  Fig S16E: Percentage of time spent freezing in mice trained in the spaced cFC task with ISI=2d, in which EBNs were stimulated via optogenetics just before Re-Training. Each row pair (A-B; D-E) is a repeated longitudinal measure from one individual mouse. Experimental group described in row 1, Training session described in row 2. Column A: Controls, freezing at Re-Training Column B: Controls, freezing at Recall Column D: EBNs CNO, freezing at Re-Training Column E: EBNs CNO, freezing at Recall Fig S16F: Percentage of time spent freezing in mice trained in the spaced cFC task with ISI=2d, in which EBNs or LBNs were stimulated during Re-Training. Each row pair (A-B; D-E; G-H) is a repeated longitudinal measure from one individual mouse. Experimental group described in row 1, Training session described in row 2. Column A: Controls, freezing at Re-Training Column B: Controls, freezing at Recall Column D: EBNs CNO, freezing at Re-Training Column E: EBNs CNO, freezing at Recall Column G: LBNs CNO, freezing at Re-Training Column H: LBNs CNO, freezing at Recall Fig S16G: Percentage of time spent freezing in mice trained in the spaced cFC task with ISI=3d, in which EBNs or LBNs were stimulated during Re-Training. Each row pair (A-B; D-E; G-H) is a repeated longitudinal measure from one individual mouse. Experimental group described in row 1, Training session described in row 2. Column A: Controls, freezing at Re-Training Column B: Controls, freezing at Recall Column D: EBNs CNO, freezing at Re-Training Column E: EBNs CNO, freezing at Recall Column G: LBNs CNO, freezing at Re-Training Column H: LBNs CNO, freezing at Recall Fig S16H: Percentage of time spent freezing in mice trained in the spaced cFC task with ISI=4d, in which EBNs or LBNs were stimulated during Re-Training. Each row pair (A-B; D-E; G-H) is a repeated longitudinal measure from one individual mouse. Experimental group described in row 1, Training session described in row 2. Column A: Controls, freezing at Re-Training Column B: Controls, freezing at Recall Column D: EBNs CNO, freezing at Re-Training Column E: EBNs CNO, freezing at Recall Column G: LBNs CNO, freezing at Re-Training Column H: LBNs CNO, freezing at Recall **adk0997_FigureS17.xls**: data underlying Fig. S17 in the main text. This file contains thirteen tabs. Fig S17B: Percentage of time spent freezing at Recall in mice trained in the so-tFC task at different ISI. Behavioral protocol described in row 1. Column A: Unpaired Column B: ISI=1d Column C: ISI=2d Column D: ISI=3d Column E: ISI=4d Fig S17D: Percentage of time spent freezing at Recall in mice trained in the so-tFC task with ISI=2d, in which EBNs were stimulated during so-Acq. Experimental group described in row 1. Column A: Controls Column B: EBNs CNO Fig S17E: Percentage of time spent freezing at Recall in mice trained in the so-tFC task with ISI=3d, in which EBNs or LBNs were stimulated during so-Acq. Experimental group described in row 1. Column A: Controls Column B: EBNs CNO Column C: LBNs CNO Fig S17F: Percentage of time spent freezing at Recall in mice trained in the so-tFC task with ISI=4d, in which LBNs were stimulated during so-Acq. Experimental group described in row 1. Column A: Controls Column B: LBNs CNO Fig S17G left: Percentage of time spent freezing before the presentation of the first CS and upon the presentation of every fo-CS (1-6) thereafter during the so-Acq session of the so-tFC task for ISI=2d, in animals where EBNs were activated during the session. Each column is a mouse. Behavioral epochs described in columns A, experimental group in row 1. Columns B-F: Controls Columns H-K: EBNs CNO Fig S17G center: Percentage of time spent freezing before the presentation of the first CS and upon the presentation of every fo-CS (1-6) thereafter during the so-Acq session of the so-tFC task for ISI=3d, in animals where EBNs or LBNs were stimulated during the session. Each column is a mouse. Behavioral epochs described in columns A, experimental group in row 1. Columns B-F: Controls Columns H-K: EBNs CNO Columns M-P: LBNs CNO  Fig S17G right: Percentage of time spent freezing before the presentation of the first CS and upon the presentation of every fo-CS (1-6) thereafter during the so-Acq session of the so-tFC task for ISI=4d, in animals where LBNs were stimulated during the session. Each column is a mouse. Behavioral epochs described in columns A, experimental group in row 1. Columns B-F: Controls Columns H-K: LBNs CNO Fig S17H left: Percentage of time spent freezing before the presentation of the first CS and upon the presentation of every so-CS (1-6) thereafter during the so-Acq session of the so-tFC task for ISI=2d, in animals where EBNs were stimulated during the session. Each column is a mouse. Behavioral epochs described in columns A, experimental group in row 1. Columns B-F: Controls Columns H-K: EBNs CNO Fig S17H center: Percentage of time spent freezing before the presentation of the first CS and upon the presentation of every so-CS (1-6) thereafter during the so-Acq session of the so-tFC task for ISI=3d, in animals where EBNs or LBNs were stimulated during the session. Each column is a mouse. Behavioral epochs described in columns A, experimental group in row 1. Columns B-F: Controls Columns H-K: EBNs CNO Columns M-P: LBNs CNO Fig S17H right: Percentage of time spent freezing before the presentation of the first CS and upon the presentation of every so-CS (1-6) thereafter during the so-Acq session of the so-tFC task for ISI=4d, in animals where LBNs were stimulated during the session. Each column is a mouse. Behavioral epochs described in columns A, experimental group in row 1. Columns B-F: Controls Columns H-K: LBNs CNO Fig S17I left: Percentage of time spent freezing before the presentation of the first CS and upon the presentation of every so-CS (1-6) thereafter during the Recall of the so-tFC task for ISI=2d, in animals where EBNs were stimulated during the so-Acq session. Each column is a mouse. Behavioral epochs described in columns A, experimental group in row 1. Columns B-F: Controls Columns H-K: EBNs CNO Fig S17I center: Percentage of time spent freezing before the presentation of the first CS and upon the presentation of every so-CS (1-6) thereafter during the Recall of the so-tFC task for ISI=3d, in animals where EBNs or LBNs were stimulated during the so-Acq session. Each column is a mouse. Behavioral epochs described in columns A, experimental group in row 1. Columns B-F: Controls Columns H-K: EBNs CNO Columns M-P: LBNs CNO Fig S17I right: Percentage of time spent freezing before the presentation of the first CS and upon the presentation of every so-CS (1-6) thereafter during the Recall of the so-tFC task for ISI=4d, in animals where LBNs were stimulated during the so-Acq session. Each column is a mouse. Behavioral epochs described in columns A, experimental group in row 1. Columns B-F: Controls Columns H-K: LBNs CNO DON008361.mat, DON008362.mat, DON008363.mat, DON010769.mat, DON010771.mat, DON011187mat, DON013198.mat, DON013199.mat, DON013223.mat, DON013388.mat, DON013426.mat, DON013885.mat, DON014305.mat, DON014306.mat, DON016320.mat, DON016357.mat, DON016358.mat, DON016360.mat, DON016361.mat, DON016883.mat, DON017272.mat: **1-photon recordings from individual mice recorded during the Acquisition, Recall 1d, and Recall 14d of a cFC or Context Only memory. Mouse ID is the name of the file, for a complete description of the recording parameters, genotype, hardware used, and behavioral protocol the animal went through see Table S1 of the original manuscript. Each file contains two different variables: dFF: The dataset comprises a matrix where each element represents the delta F over F (dF/F0) value of the fluorescence signal recorded from neurons over time. Specifically, each column in the matrix corresponds to an individual neuron, while each row represents a distinct frame of the recording. The number of rows in the matrix equals the number of frames, reflecting the temporal resolution of the recording. Data from the three recording sessions were concatenated (see description of the second variable, nFrSes). The number of columns corresponds to the number of neurons that were recorded and longitudinally tracked in each mouse. The value in each cell of this matrix indicates the dF/F0 for a particular neuron at a given frame, expressed in arbitrary units. The dF/F0 value is calculated as the change in fluorescence intensity relative to a baseline fluorescence level, providing a measure of the relative change in fluorescence over time. If no calcium event was recoded for a longitudinally tracked ROI during an experimental session, the corresponding cell is NaN. nFrSes: describes the number of frames corresponding to different recording epochs across various experimental days. In this matrix, each row represents a specific recording context, while each column represents a different experimental day. The rows are organized such that the first row corresponds to recordings made in the Home Cage, and the second row corresponds to recordings made in the Training Context. The columns are designated for different experimental days: Acquisition, Recall 1d, and Recall 14d.