Fig. 2

Knockout of PUMA elevates Rad51 in PSCs or HPCs. a Immunoblotting (IB) of Rad51, 53BP1, and PUMA expression in PUMA WT and KO PSCs or HPCs after IR. PSCs and HPCs were treated with 2 Gy IR, and then collected at 0 or 8 h post-IR for IB analysis. Actin was used as a control. b Quantification of positive cells with Rad51 foci in PUMA WT and KO PSCs at indicated time points after IR. c Representative images of Rad51 foci in PUMA WT and KO PSCs after IR. Scale bars, 10 μm. d Effects of PUMA re-expression on expression of Rad51 and 53BP1 in PUMA KO PSCs after IR. Lentivirus-mediated PUMA was infected into PUMA KO PSCs, and then selected the clones of transfected cells expressing a similar level of PUMA as that PUMA WT cells. e Effect of re-expression of PUMA on HR repair in PUMA KO PSCs. f IB of expression of Rad51 and 53BP1 in PSCs or HPCs with a Rad51 shRNA. g Effect of Rad51 knockdown on PUMA-mediated HR repair. Data are representative of three independent experiments with similar results. Error bars, SD. *P < 0.05, **P < 0.01, ***P < 0.001