DOI: 10.5061/dryad.rxwdbrvnt
title: Uncropped gel—nuclease data from: A nicking class 1 OLD family nuclease encoded by <em>Vibrio cholerae</em> inhibits virbiophage replication and is countered by a direct inhibitor

description: Bacteria are constantly threatened by their viral predators (phages),
 which has resulted in the development of defense systems for bacterial
 survival. One family of defense systems found widely across bacteria are
 OLD (for overcoming lysogenization defect) family nucleases. Despite
 recent discoveries regarding Class 2 and 4 OLD family nucleases and how
 phages overcome them, Class 1 OLD family nucleases warrant further study,
 as there has only been one anti-phage Class 1 OLD family nuclease
 described to date. Here, we identify the first bacteria-encoded Class 1
 OLD family nuclease that exerts anti-phage activity: the Vibrio cholerae
 encoded Class 1 OLD family nuclease Vc OLD.  We describe its
 disruption of the genome replication of the lytic vibriophage ICP1.
 Furthermore, we examine its in vitro activity, identifying Vc OLD as a DNA
 nickase. Finally, we identify the first direct inhibitor of a Class 1 OLD
 family nuclease, the ICP1-encoded Oad1. Our research further illuminates
 Class 1 OLD family nucleases’ role in phage defense and explores the
 dynamic arms race between V. cholerae and its predatory phage ICP1.
description: Nuclease assays with purified Vc OLD and Oad1 and bovine serum
 albumin (BSA) (Fisher BP1600) were carried out in 50-mM Tris–HCl,
 pH = 7.4, 50-mM NaCl, 10-mM MgCl<sub>2</sub>, 10-mM
 MnCl<sub>2</sub>, 10-mM CaCl<sub>2</sub>,
 10NiCl<sub>2</sub>, and 10-mM ZnCl<sub>2</sub>.
 Frozen samples of all proteins were thawed and added to the buffer at
 varying concentrations. All nucleic acid substrates were eluted into
 H<sub>2</sub>O after nucleic acid collection. Double-stranded
 DNA (dsDNA) substrate was extracted from bacteria or phage following the
 procedure outlined in “Post-infection deep sequencing and coverage
 mapping.” Similarly, single-stranded DNA (ssDNA) (from M13 phage) was
 ordered from New England Biolabs (N4040S), extracted, and eluted into
 water. Total RNA was prepped from <em>V. cholerae</em> E7946
 as described previously. Briefly, Bacterial strains were grown to an
 OD<sub>600</sub> = 0.3, collected, and mixed 1:1 with ice-cold
 methanol. Methanol-treated samples were pelleted at 7000 ×
 <em>g</em> at 4°C, and the resulting pellets were washed in
 ice-cold 1× phosphate-buffered saline. The washed pellets were resuspended
 in 200 μl TRI Reagent (Millipore/Sigma) and incubated for 5 min at room
 temperature. The samples were mixed with 40 μl chloroform, vortexed, and
 incubated for 10 min at room temperature. The chloroform-treated samples
 were then centrifuged at 12 000 × <em>g</em> for 10 min at
 4°C. Following centrifugation, the upper (aqueous) phase was collected,
 promptly mixed with 110 μl 2-propanol and 11 μl pH 6.2 3-M sodium acetate,
 and mixed vigorously. The samples were centrifuged at 12 000 ×
 <em>g</em> for 15 min at 4°C, and the pellets were washed
 twice with 75% ethanol. The washed pellets were incubated at 65°C for 3
 min to evaporate residual ethanol. The RNA was resuspended in 10–30 μl of
 diethyl dicarbonate water, and RNA concentration were analyzed by
 NanoDrop. Plasmid DNA substrate, a pUC19 derivative, was prepped using the
 NEB Monarch Plasmid Miniprep Kit. All DNA substrates were added at 100 ng
 per 20 μl reaction. RNA was loaded at 500 ng per 20 μl reaction. Reactions
 containing DNA substrate were incubated at 37°C for 3 h, and then 1 μg/ml
 of Proteinase K was added to each reaction to digest bound protein and
 incubated at 37°C for 30 min. Reactions containing RNA substrate were
 incubated at 37°C for 30 min, and then 1 μg/ml of Proteinase K was added
 to each reaction to digest bound protein and incubated at 37°C for 30 min.
 To generate linear or nicked controls, 100 ng of vector was digested in 1×
 CutSmart Buffer for 1 h at 37°C with 10 U of BamHI-HF (NEB) or 10 U of
 Nb.BtsI (NEB) in 20 μl reactions, respectively, and then 1 μg/ml of
 Proteinase K was added to each reaction to digest bound protein and
 incubated at 37°C for 30 min. The entire reaction volume was loaded and
 run on a 0.8% agarose gel stained with GelRed.
description: # Uncropped gel—nuclease data from: A nicking class 1 OLD family nuclease
 encoded by *Vibrio cholerae* inhibits virbiophage replication and is
 countered by a direct inhibitor Dataset DOI:
 [10.5061/dryad.rxwdbrvnt](10.5061/dryad.rxwdbrvnt) ## Description of the
 data and file structure ### Files and variables #### File:
 Figure_S15A-_Replicate_B.tif **Description:** These data are linked to
 Supplemental Figure 15 of the manuscript. Uncropped image of a 0.8%
 agarose gel. 20uL of rxn was loaded per lane. From left to right, the
 lanes are: Plasmid incubated with BamHI-HF (NEB), Plasmid nicked with
 Nb.BtsI (NEB), plasmid not incubated with enzyme, plasmid incubated with
 100nM Vc OLD and 2nM BSA, plasmid incubated with 100nM Vc OLD and 4nM BSA,
 plasmid incubated with 100nM Vc OLD and 100nM BSA, plasmid incubated with
 100nM Vc OLD and 2500nM BSA, plasmid incubated with 100nM Vc OLD and
 5000nM BSA, plasmid incubated with 5000nM BSA and without Vc OLD, 100nM Vc
 OLD incubated without plasmid, and 5000nM BSA incubated without plasmid.
 Replicate B. #### File: Figure_S15A-_Replicate_C.tif
 **Description:** These data are linked to Supplemental Figure 15 of the
 manuscript. Uncropped image of a 0.8% agarose gel. 20uL of rxn was loaded
 per lane. From left to right, the lanes are: Plasmid incubated with
 BamHI-HF (NEB), Plasmid nicked with Nb.BtsI (NEB), plasmid not incubated
 with enzyme, plasmid incubated with 100nM Vc OLD and 2nM BSA, plasmid
 incubated with 100nM Vc OLD and 4nM BSA, plasmid incubated with 100nM Vc
 OLD and 100nM BSA, plasmid incubated with 100nM Vc OLD and 2500nM BSA,
 plasmid incubated with 100nM Vc OLD and 5000nM BSA, plasmid incubated with
 5000nM BSA and without Vc OLD, 100nM Vc OLD incubated without plasmid, and
 5000nM BSA incubated without plasmid. Replicate C. #### File:
 Figure_S15C-_Replicate_A.tif **Description:** These data are linked to
 Supplemental Figure 15 of the manuscript. Uncropped image of a 0.8%
 agarose gel. 20uL of rxn was loaded per lane. From left to right, the
 lanes are: Plasmid incubated with BamHI-HF (NEB), Plasmid nicked with
 Nb.BtsI (NEB), plasmid not incubated with enzyme, plasmid incubated with
 100nM Vc OLD and 2nM OAD1, plasmid incubated with 100nM Vc OLD and 4nM
 OAD1, plasmid incubated with 100nM Vc OLD and 100nM OAD1, plasmid
 incubated with 100nM Vc OLD and 2500nM OAD1, plasmid incubated with 100nM
 Vc OLD and 5000nM OAD1, plasmid incubated with 5000nM OAD1 and without Vc
 OLD, 100nM Vc OLD incubated without plasmid, and 5000nM OAD1 incubated
 without plasmid. Replicate A. #### File: Figure_S15A-_Replicate_A.tif
 **Description:** These data are linked to Supplemental Figure 15 of the
 manuscript. Uncropped image of a 0.8% agarose gel. 20uL of rxn was loaded
 per lane. From left to right the lanes are: Plasmid incubated with
 BamHI-HF (NEB) , Plasmid nicked with Nb.BtsI (NEB), plasmid not incubated
 with enzyme, plasmid incubated with 100nM Vc OLD and 2nM BSA, plasmid
 incubated with 100nM Vc OLD and 4nM BSA, plasmid incubated with 100nM Vc
 OLD and 100nM BSA, plasmid incubated with 100nM Vc OLD and 2500nM BSA,
 plasmid incubated with 100nM Vc OLD and 5000nM BSA, plasmid incubated with
 5000nM BSA and without Vc OLD, 100nM Vc OLD incubated without plasmid, and
 5000nM BSA incubated without plasmid. Replicate A. #### File:
 Figure_S15C-_Replicate_C.tif **Description:** These data are linked to
 Supplemental Figure 15 of the manuscript. Uncropped image of a 0.8%
 agarose gel. 20uL of rxn was loaded per lane. From left to right, the
 lanes are: Plasmid incubated with BamHI-HF (NEB), Plasmid nicked with
 Nb.BtsI (NEB), plasmid not incubated with enzyme, plasmid incubated with
 100nM Vc OLD and 2nM OAD1, plasmid incubated with 100nM Vc OLD and 4nM
 OAD1, plasmid incubated with 100nM Vc OLD and 100nM OAD1, plasmid
 incubated with 100nM Vc OLD and 2500nM OAD1, plasmid incubated with 100nM
 Vc OLD and 5000nM OAD1, plasmid incubated with 5000nM OAD1 and without Vc
 OLD, 100nM Vc OLD incubated without plasmid, and 5000nM OAD1 incubated
 without plasmid. Replicate C. #### File: Figure_S15C-_Replicate_B.tif
 **Description:** These data are linked to Supplemental Figure 15 of the
 manuscript. Uncropped image of a 0.8% agarose gel. 20uL of rxn was loaded
 per lane. From left to right, the lanes are: Plasmid incubated with
 BamHI-HF (NEB), Plasmid nicked with Nb.BtsI (NEB), plasmid not incubated
 with enzyme, plasmid incubated with 100nM Vc OLD and 2nM OAD1, plasmid
 incubated with 100nM Vc OLD and 4nM OAD1, plasmid incubated with 100nM Vc
 OLD and 100nM OAD1, plasmid incubated with 100nM Vc OLD and 2500nM OAD1,
 plasmid incubated with 100nM Vc OLD and 5000nM OAD1, plasmid incubated
 with 5000nM OAD1 and without Vc OLD, 100nM Vc OLD incubated without
 plasmid, and 5000nM OAD1 incubated without plasmid. Replicate B. ####
 File: Figure_S9C-_Replicate_C.tif **Description:** These data are linked
 to Supplemental Figure 9 of the manuscript. Uncropped image of a 0.8%
 agarose gel. 20uL of rxn was loaded per lane. For all of these rxns, the
 plasmid substrate contained 41 bp from the *V. cholerae* genome. From left
 to right the lanes are: a DNA ladder, Plasmid incubated with BamHI-HF
 (here the plasmid is not linearized due to the loss of the BamHI-HF
 cutting site), Plasmid nicked with Nb.BtsI (NEB), plasmid not incubated
 with enzyme, plasmid incubated with 500nM Vc OLD, plasmid incubated with
 250nM Vc OLD, plasmid incubated with 100nM Vc OLD, plasmid incubated with
 50nM Vc OLD, plasmid incubated with 25nM Vc OLD, plasmid incubated with
 10nM Vc OLD, and 500nM Vc OLD incubated without plasmid. Replicate C. ####
 File: Figure_S9A-_Replicate_A.tif **Description:** These data are linked
 to Supplemental Figure 9 of the manuscript. Uncropped image of a 0.8%
 agarose gel. 20uL of rxn was loaded per lane. For all of these rxns, the
 plasmid substrate contained 41 bp from ICP1 genome. From left to right the
 lanes are: a DNA ladder, Plasmid incubated with BamHI-HF (here the plasmid
 is not linearized due to the loss of the BamHI-HF cutting site), Plasmid
 nicked with Nb.BtsI (NEB), plasmid not incubated with enzyme, plasmid
 incubated with 500nM Vc OLD, plasmid incubated with 250nM Vc OLD, plasmid
 incubated with 100nM Vc OLD, plasmid incubated with 50nM Vc OLD, plasmid
 incubated with 25nM Vc OLD, plasmid incubated with 10nM Vc OLD, and 500nM
 Vc OLD incubated without plasmid. Replicate A. #### File:
 Figure_S9C-_Replicate_B.tif **Description:** These data are linked to
 Supplemental Figure 9 of the manuscript. Uncropped image of a 0.8% agarose
 gel. 20uL of rxn was loaded per lane. For all of these rxns, the plasmid
 substrate contained 41 bp from the *V. cholerae* genome. From left to
 right the lanes are: a DNA ladder, Plasmid incubated with BamHI-HF (here
 the plasmid is not linearized due to the loss of the BamHI-HF cutting
 site), Plasmid nicked with Nb.BtsI (NEB), plasmid not incubated with
 enzyme, plasmid incubated with 500nM Vc OLD, plasmid incubated with 250nM
 Vc OLD, plasmid incubated with 100nM Vc OLD, plasmid incubated with 50nM
 Vc OLD, plasmid incubated with 25nM Vc OLD, plasmid incubated with 10nM Vc
 OLD, and 500nM Vc OLD incubated without plasmid. Replicate B. #### File:
 Figure_S9A-_Replicate_C.tif **Description:**  These data are linked to
 Supplemental Figure 9 of the manuscript. Uncropped image of a 0.8% agarose
 gel. 20uL of rxn was loaded per lane. For all of these rxns, the plasmid
 substrate contained 41 bp from ICP1 genome. From left to right the lanes
 are: a DNA ladder, Plasmid incubated with BamHI-HF (here the plasmid is
 not linearized due to the loss of the BamHI-HF cutting site), Plasmid
 nicked with Nb.BtsI (NEB), plasmid not incubated with enzyme, plasmid
 incubated with 500nM Vc OLD, plasmid incubated with 250nM Vc OLD, plasmid
 incubated with 100nM Vc OLD, plasmid incubated with 50nM Vc OLD, plasmid
 incubated with 25nM Vc OLD, plasmid incubated with 10nM Vc OLD, and 500nM
 Vc OLD incubated without plasmid. Replicate C. #### File:
 Figure_S9C-_Replicate_A.tif **Description:**  These data are linked to
 Supplemental Figure 9 of the manuscript. Uncropped image of a 0.8% agarose
 gel. 20uL of rxn was loaded per lane. For all of these rxns, the plasmid
 substrate contained 41 bp from the *V. cholerae* genome. From left to
 right the lanes are: a DNA ladder, Plasmid incubated with BamHI-HF (here
 the plasmid is not linearized due to the loss of the BamHI-HF cutting
 site), Plasmid nicked with Nb.BtsI (NEB), plasmid not incubated with
 enzyme, plasmid incubated with 500nM Vc OLD, plasmid incubated with 250nM
 Vc OLD, plasmid incubated with 100nM Vc OLD, plasmid incubated with 50nM
 Vc OLD, plasmid incubated with 25nM Vc OLD, plasmid incubated with 10nM Vc
 OLD, and 500nM Vc OLD incubated without plasmid. Replicate A. #### File:
 Figure_S9A-_Replicate_B.tif **Description:**  These data are linked to
 Supplemental Figure 9 of the manuscript. Uncropped image of a 0.8% agarose
 gel. 20uL of rxn was loaded per lane. For all of these rxns, the plasmid
 substrate contained 41 bp from ICP1 genome. From left to right the lanes
 are: a DNA ladder, Plasmid incubated with BamHI-HF (here the plasmid is
 not linearized due to the loss of the BamHI-HF cutting site), Plasmid
 nicked with Nb.BtsI (NEB), plasmid not incubated with enzyme, plasmid
 incubated with 500nM Vc OLD, plasmid incubated with 250nM Vc OLD, plasmid
 incubated with 100nM Vc OLD, plasmid incubated with 50nM Vc OLD, plasmid
 incubated with 25nM Vc OLD, plasmid incubated with 10nM Vc OLD, and 500nM
 Vc OLD incubated without plasmid. Replicate B. #### File:
 Figure_4_-_Replicate_C.tif **Description:** These data are linked to
 Figure 4 of the manuscript. Uncropped image of a 0.8% agarose gel. 20uL of
 rxn was loaded per lane. From left to right the lanes are: Plasmid
 linearized with BamHI-HF (NEB), Plasmid nicked with Nb.BtsI (NEB), plasmid
 not incubated with enzyme, plasmid incubated with 500nM Vc OLD, plasmid
 incubated with 250nM Vc OLD, plasmid incubated with 100nM Vc OLD, plasmid
 incubated with 50nM Vc OLD, plasmid incubated with 25nM Vc OLD, plasmid
 incubated with 10nM Vc OLD, and 500nM Vc OLD incubated without plasmid.
 Replicate C. #### File: Figure_4_-_Replicate_B.tif **Description:** These
 data are linked to Figure 4 of the manuscript. Uncropped image of a 0.8%
 agarose gel. 20uL of rxn was loaded per lane. From left to right the lanes
 are: Plasmid linearized with BamHI-HF (NEB), Plasmid nicked with Nb.BtsI
 (NEB), plasmid not incubated with enzyme, plasmid incubated with 500nM Vc
 OLD, plasmid incubated with 250nM Vc OLD, plasmid incubated with 100nM Vc
 OLD, plasmid incubated with 50nM Vc OLD, plasmid incubated with 25nM Vc
 OLD, plasmid incubated with 10nM Vc OLD, and 500nM Vc OLD incubated
 without plasmid. Replicate B. #### File: Figure_4_-_Replicate_A.tif
 **Description:** These data are linked to Figure 4 of the manuscript.
 Uncropped image of a 0.8% agarose gel. 20uL of rxn was loaded per lane.
 From left to right the lanes are: Plasmid linearized with BamHI-HF (NEB),
 Plasmid nicked with Nb.BtsI (NEB), plasmid not incubated with enzyme,
 plasmid incubated with 500nM Vc OLD, plasmid incubated with 250nM Vc OLD,
 plasmid incubated with 100nM Vc OLD, plasmid incubated with 50nM Vc OLD,
 plasmid incubated with 25nM Vc OLD, plasmid incubated with 10nM Vc OLD,
 and 500nM Vc OLD incubated without plasmid. Replicate A. #### File:
 Figure_S8-_Replicate_B.tif **Description:** These data are linked to
 Supplementary Figure 8 of the manuscript. Uncropped image of a 0.8%
 agarose gel. 20uL of rxn was loaded per lane. From left to right the lanes
 are: Plasmid linearized with BamHI-HF (NEB) without ATP, Plasmid nicked
 with Nb.BtsI (NEB) without ATP, plasmid not incubated with enzyme or ATP,
 plasmid incubated with 100nM Vc OLD and 0.05mM ATP, plasmid incubated with
 100nM Vc OLD and 0.1mM ATP, plasmid incubated with 100nM Vc OLD and and
 0.2mM ATP, plasmid incubated with 100nM Vc OLD and and 0.5mM ATP, plasmid
 incubated with 100nM Vc OLD and and 1.0mM ATP, plasmid incubated without
 enzyme and with 1.0mM ATP 10nM Vc OLD, and 100nM Vc OLD incubated without
 plasmid or ATP. Replicate B. #### File: Figure_S8-_Replicate_C.tif
 **Description:** These data are linked to Supplementary Figure 8 of the
 manuscript. Uncropped image of a 0.8% agarose gel. 20uL of rxn was loaded
 per lane. From left to right the lanes are: Plasmid linearized with
 BamHI-HF (NEB) without ATP, Plasmid nicked with Nb.BtsI (NEB) without ATP,
 plasmid not incubated with enzyme or ATP, plasmid incubated with 100nM Vc
 OLD and 0.05mM ATP, plasmid incubated with 100nM Vc OLD and 0.1mM ATP,
 plasmid incubated with 100nM Vc OLD and and 0.2mM ATP, plasmid incubated
 with 100nM Vc OLD and and 0.5mM ATP, plasmid incubated with 100nM Vc OLD
 and and 1.0mM ATP, plasmid incubated without enzyme and with 1.0mM ATP
 10nM Vc OLD, and 100nM Vc OLD incubated without plasmid or ATP. Replicate
 C. #### File: Figure_S8-_Replicate_A.tif **Description:** These data are
 linked to Supplementary Figure 8 of the manuscript. Uncropped image of a
 0.8% agarose gel. 20uL of rxn was loaded per lane. From left to right the
 lanes are: Plasmid linearized with BamHI-HF (NEB) without ATP, Plasmid
 nicked with Nb.BtsI (NEB) without ATP, plasmid not incubated with enzyme
 or ATP, plasmid incubated with 100nM Vc OLD and 0.05mM ATP, plasmid
 incubated with 100nM Vc OLD and 0.1mM ATP, plasmid incubated with 100nM Vc
 OLD and and 0.2mM ATP, plasmid incubated with 100nM Vc OLD and and 0.5mM
 ATP, plasmid incubated with 100nM Vc OLD and and 1.0mM ATP, plasmid
 incubated without enzyme and with 1.0mM ATP 10nM Vc OLD, and 100nM Vc OLD
 incubated without plasmid or ATP. Replicate A. #### File:
 Figure_S7C_-_Replicates_A_and_B.tif **Description:** These data are linked
 to Supplementary Figure 7C of the manuscript. Uncropped image of a 0.8%
 agarose gel. 20uL of rxn was loaded per lane. From left to right, lanes
 contain: a DNA ladder, bacterial RNA incubated with RNAse, bacterial RNA
 not incubated with enzyme, bacterial RNA incubated with 500nM Vc OLD,
 500nM Vc OLD incubated without bacterial RNA, a DNA ladder, bacterial RNA
 incubated with RNAse, bacterial RNA not incubated with enzyme, bacterial
 RNA incubated with 500nM Vc OLD, and 500nM Vc OLD incubated without
 bacterial RNA. Each five lanes represent an independent replicate. ####
 File: Figure_S7A_-_Replicate_C.tif **Description:** These data are linked
 to Supplementary Figure 7A of the manuscript. Uncropped image of a 0.8%
 agarose gel. 20uL of rxn was loaded per lane. From left to right, lanes
 contain: a DNA ladder, phage gDNA incubated with DNAse, phage gDNA not
 incubated with enzyme, phage gDNA incubated with 500nM Vc OLD, 500nM Vc
 OLD incubated without gDNA, bacterial gDNA incubated with DNAse, bacterial
 gDNA not incubated with enzyme, bacterial gDNA incubated with 500nM Vc
 OLD, 500nM Vc OLD incubated without gDNA, phage gDNA incubated with DNAse,
 phage gDNA not incubated with enzyme, and phage gDNA incubated with 500nM
 Vc OLD. The lanes two through nine are the third replicates of the
 experiment. The final three lanes are the second replicate of this
 experiment with phage gDNA. #### File:
 Figure_S7A-*plasmid*-_Replicate_A-C.tif **Description:** These data are
 linked to Supplementary Figure 7A of the manuscript. Uncropped image of a
 0.8% agarose gel. 20uL of rxn was loaded per lane. Left to right, lanes
 contain: plasmid incubated with DNAse, plasmid gDNA not incubated with
 enzyme, plasmid gDNA incubated with 500nM Vc OLD, 500nM Vc OLD incubated
 without gDNA, plasmid incubated with DNAse, plasmid gDNA not incubated
 with enzyme, plasmid gDNA incubated with 500nM Vc OLD, 500nM Vc OLD
 incubated without gDNA, plasmid incubated with DNAse, plasmid gDNA not
 incubated with enzyme, plasmid gDNA incubated with 500nM Vc OLD, and 500nM
 Vc OLD incubated without gDNA. Each four lanes is a replicate of this
 experiment. #### File: Figure_S7B_-_Replicate_A-C.tif
 **Description:** These data are linked to Supplementary Figure 7B of the
 manuscript. Uncropped image of a 0.8% agarose gel. 20uL of rxn was loaded
 per lane. Left to right, lanes contain: ssDNA incubated with DNAse, ssDNA
 not incubated with enzyme, ssDNA incubated with 500nM Vc OLD, 500nM Vc OLD
 incubated without ssDNA, ssDNA incubated without enzyme, ssDNA incubated
 with 500nM Vc OLD, ssDNA incubated with DNAse, 500nM Vc OLD incubated
 without ssDNA, ssDNA incubated with DNAse, ssDNA not incubated with
 enzyme, ssDNA incubated with 500nM Vc OLD, and 500nM Vc OLD incubated
 without ssDNA. Each four lanes is a replicate of this experiment. ####
 File: Figure_S7A_-_Replicate_B.tif **Description:** These data are linked
 to Supplementary Figure 7A of the manuscript. Uncropped image of a 0.8%
 agarose gel. 20uL of rxn was loaded per lane. Red boxes indicates lanes
 not applicable to this study. For the lanes applicable to this study, left
 to right contain: phage gDNA incubated with DNAse, phage gDNA not
 incubated with enzyme, phage gDNA incubated with 500nM Vc OLD, 500nM Vc
 OLD incubated without gDNA, bacterial gDNA incubated with DNAse, bacterial
 gDNA not incubated with enzyme, bacterial gDNA incubated with 500nM Vc
 OLD, and 500nM Vc OLD incubated without gDNA. All of these are the second
 replicates of the experiment. #### File:
 Figure_S7A_-_Bacterial_gDNA_Replicate.tif **Description:** These data are
 linked to Supplementary Figure 7A of the manuscript. Uncropped image of a
 0.8% agarose gel. 20uL of rxn was loaded per lane. Red boxes indicates
 lanes not applicable to this study. For the lanes applicable to this study
 (middle), left to right contain: bacterial gDNA incubated with DNAse,
 bacterial gDNA not incubated with enzyme, bacterial gDNA incubated with
 500nM Vc OLD, and 500nM Vc OLD incubated without gDNA. #### File:
 Figure_S7C_-_Replicates_C.tif **Description:** These data are linked to
 Supplementary Figure 7C of the manuscript. Uncropped image of a 0.8%
 agarose gel. 20uL of rxn was loaded per lane. The red box indicates lanes
 not applicable to this study. For lanes applicable, from left to right,
 lanes contain: a DNA ladder, bacterial RNA incubated with RNAse, bacterial
 RNA not incubated with enzyme, bacterial RNA incubated with 500nM Vc OLD,
 and 500nM Vc OLD incubated without bacterial RNA. This is the third
 replicate of this experiment. ## Code/software N/A ## Access information
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