Fig. 4

The HR defect caused by BRCA1-depletion is not further affected by PARG depletion. a and b The frequency of HR-mediated repair events was analysed by flow cytometry in U2OS-DR-GFP-mCherry-I-SceI-GR cells after transfection with the indicated siRNA and induction of DSB formation by incubation with TA for 48 h. The percentage of Cherry- and GFP-positive cells is indicated. In b, the values correspond to the ratio of GFP-positive cells relative to cells transfected with scrambled siRNA (siCTL), as illustrated in a, and represent the mean ± SD of 7 independent experiments. c and d Defect of etoposide-induced RAD51 foci formation in BRCA1-depleted cells is not further affected by PARG depletion. Cells transfected with the indicated siRNA were incubated with 10 µM etoposide for 1 h, released into fresh medium for 2 h and processed for immunofluorescence using anti-RAD51 and anti-γH2AX antibodies. Representative immunofluorescence image are shown in c. In d, the box plot graph depicts the percentage of cells displaying RAD51 foci (more than 10 RAD51 foci per cell) from 5 independent experiments scoring >200 nuclei for each condition. p: 0.01 < * < 0.05; 0.05 < ns, not significant