Fig. 4: ZNF827 promotes HR-directed DNA repair. | Nature Communications

Fig. 4: ZNF827 promotes HR-directed DNA repair.

From: ZNF827 is a single-stranded DNA binding protein that regulates the ATR-CHK1 DNA damage response pathway

Fig. 4

A Representative images of sister-chromatid exchange (SCE) events in U-2 OS and U-2 OS ZNF827 CRISPR KO cells incubated with topotecan or DMSO. Green boxes indicate sites of exchange events. Scale bar represents 5 μm. B Quantitative data of SCE events in U-2 OS and U-2 OS ZNF827 CRISPR KO cells incubated with topotecan or DMSO (left panel), and in HT1080 cells following 72 h ZNF827 depletion, including treatment with topotecan or DMSO (right panel). Data presented as mean + SEM from 800–1000 chromosomes per replicate from three biological replicates; ****P < 0.0001 by one-way ANOVA C HR reporter assay in U-2 OS cells stably expressing pDR-GFP. Quantitation of GFP-positive cells detected by flow cytometry. Data presented as mean + SEM from three independent experiments; ****P < 0.0001 by two-tailed t-test. D Representative images of BRCA1 (green) and pRPA32 (S33) (red) foci in HT1080 cells. Scale bar represents 5 μm. Quantitation of E BRCA1 colocalizations with pRPA32 (S33) per nucleus, and F BRCA1 foci per nucleus in HT1080 cells. Out of three experiments, n = 930 cells scored in scrambled DMSO, n = 694 cells scored in siZNF827 DMSO, n = 1054 cells scored in scrambled topotecan, and n = 458 cells scored in siZNF827 topotecan using CellProfiler. At least 25% overlap criterion was applied for colocalization count. Data are representative of three independent experiments and presented as Tukey box plots; ****P < 0.0001 by Kruskal–Wallis tests. G Growth curves by cell confluency obtained from IncuCyte® live cell assays of U-2 OS cells following ZNF827 depletion. Topotecan or DMSO were added at 24 h post-siRNA knockdown and cells imaged every 4 h up to 160 h. Data presented as mean ± SEM from two biological replicates. H Growth curves by cell confluency obtained from IncuCyte® live cell assays of HT1080 cells following ZNF827 depletion. Topotecan or DMSO were added at 24 h post-siRNA knockdown and cells imaged every 4 h up to 160 h. Data presented as mean ± SEM from two biological replicates. Source data are provided as a Source Data file.

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