DOI: 10.5061/dryad.9w0vt4bq3
title: Data from: Differences in RAD51 transcriptional response and cell cycle dynamics reveal varying sensitivity to DNA damage among Arabidopsis thaliana root cell types

description: Throughout their lifecycle, plants are subjected to DNA damage from
 various sources, both environmental and endogenous. Investigating the
 mechanisms of the DNA damage response (DDR) is essential to unravel how
 plants adapt to the changing environment, which can induce varying amounts
 of DNA damage. Using a combination of whole-mount single-molecule
 RNA fluorescence in situ hybridization (WM-smFISH) and plant cell
 cycle reporter lines, we investigated the transcriptional activation of a
 key homologous recombination (HR) gene, RAD51, in response to increasing
 amounts of DNA damage in Arabidopsis thaliana roots. The results uncover
 consistent variations in RAD51 transcriptional response and cell cycle
 arrest among distinct cell types and developmental zones. Furthermore, we
 demonstrate that DNA damage induced by genotoxic stress results in RAD51
 transcription throughout the whole cell cycle, dissociating its
 traditional link with S/G2 phases. This work advances the current
 comprehension of DNA damage response in plants by demonstrating
 quantitative differences in DDR activation. In addition, it reveals new
 associations with the cell cycle and cell types, providing crucial
 insights for further studies of the broader response mechanisms in plants.
description: # Data from: Differences in RAD51 transcriptional response and cell cycle
 dynamics reveal varying sensitivity to DNA damage among Arabidopsis
 thaliana root cell types
 [https://doi.org/10.5061/dryad.9w0vt4bq3](https://doi.org/10.5061/dryad.9w0vt4bq3) Here we provide all the raw data images for the manuscript: Kutashev et al. 2024 *New Phytologist* (DOI: 10.1111/NPH.19875). ## Description of the data and file structure Files are organized according to figure numbers as described in the manuscript (DOI: 10.1111/NPH.19875). Please refer to the methods section of the manuscript for details on the imaging settings. Figure 1 - Raw microscopy images for Immunofluorescence (anti-gammaH2AX) and single-molecule RNA FISH (detection of RAD51 transcripts). Figure 2 - Raw microscopy images for wholemount single molecule RNA FISH (detection of RAD51 transcripts). Figure 3 - Raw microscopy images for wholemount single molecule RNA FISH using RAD51-GFP line (detection of RAD51 protein and RNAs). Fugure 4- RAW microscopy images for cell cycle investigation (CDT1a-CFP; Cytrap; EdU; WM-smFISH/EdU) Fig. S1. Raw microscopy images used for evaluation of *RAD51* and *PP2A* transcription using single molecule RNA FISH Fig. S2. Raw microscopy images used for quantification of *RAD51* transcription sites and mRNAs per cell from Arabidopsis  roots using WM-smFISH. Fig. S3. Raw microscopy images used for quantification of *RAD51* mRNA molecules in Arabidopsis roots exposed to increasing amounts of DNA damage using WM-smFISH. Fig. S4. Magnified representative images from Arabidopsis root cells with different number of RAD51 mRNAs obtained by WM-smFISH. Fig. S5. Raw microscopy images used for quantification of *RAD51* mRNA molecules per cell in stem cell cells of Arabidopsis roots. Fig. S7. Raw microscopy images of *RAD51* WM-smFISH in meristematic and elongation zones of Arabidopsis roots . Fig. S8. Raw microscopy images used for quantification of *RAD51-GFP* mRNA and RAD51-GFP protein signals in roots of Arabidopsis RAD51-GFP line treated with 0 μM and 10 μM zeocin. Fig. S9. Raw microscopy images used for quantification of *RAD51-GFP* mRNA and RAD51-GFP protein signals in roots of Arabidopsis RAD51-GFP line treated with 50 μM of zeocin. Fig. S10. Raw microscopy images used for quantification of *RAD51-GFP* mRNA and RAD51-GFP protein signals in roots of Arabidopsis RAD51-GFP line treated with 170 μM of zeocin. Fig. S11. Raw microscopy images used for evaluation of cell cycle arrest in Arabidopsis roots using EdU staining. Fig. S12. Raw microscopy images used for evaluation of cell cycle changes using Cytrap line in roots after exposure different concentrations of zeocin. Fig. S13. Raw microscopy images used for quantification of DAPI signal intensity in Arabidopsis roots exposed to different concentrations of zeocin. Fig. S14. Raw microscopy images used for evaluation of cell cycle changes using PlaCCI line in roots after exposure to different concentrations of zeocin. Fig. S15. Raw microscopy images used for assessment of *RAD51* mRNA molecule half-life. Fig. S16. Raw microscopy images used for quantification of *RAD51* mRNAs cell in cells with (+) and without (-) EdU staining. Table S2. Raw microscopy images used for quantification of *RAD51* transcription sites.