Fig. 5: BRD4 isoform A rescues R-loop-induced DNA damage. | Nature Communications

Fig. 5: BRD4 isoform A rescues R-loop-induced DNA damage.

From: BRD4 prevents the accumulation of R-loops and protects against transcription–replication collision events and DNA damage

Fig. 5

a WB of endogenous levels of BRD4 isoforms A, B, and C probed with a pan-BRD4 antibody following isoform-specific siRNA knockdown. b Left panel: IF images of γH2AX and S9.6 fluorescence in HeLa cells following siRNA knockdown of BRD4 isoforms A, B, or C. Right panel: Quantification of γH2AX foci and S9.6 nuclear intensities (n = 3 separate experiments). γH2AX foci are shown as mean ± SEM. Box-whisker plots for S9.6 intensity drawn as in Fig. 1b. Significance assessed using ANOVA followed by Tukey’s test ****Adjusted P < 0.0001. Scale bar = 2.5 μm. c Left panel: IF images of γH2AX fluorescence in cells overexpressing GFP-tagged BRD4 isoforms (solid arrows) and in cells not overexpressing GFP-tagged BRD4 isoforms (open arrows) following JQ1 treatment. Right panel: quantification of γH2AX foci and integrated nuclear intensity per cell (n = 3 separate experiments) presented as mean ± SEM. Significance assessed using two-tailed unpaired t test for γH2AX foci per cell, ****P < 0.0001. Scale bar in b, c = 5 μm. Source data are provided as a Source data file.

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