Fig. 3

PUMA binds to cytosolic Rad51 and promotes its ubiquitination and stability. a Immunoprecipitation (IP) and IB of PUMA interaction with Rad51 in PSCs or HPCs. PSC and HPC cells were treated with 2 Gy of IR, and then collected at 0 or 8 h for IP and IB analyses. β-Actin was used as a loading control. b IP and IB of Rad51 ubiquitination in PSCs treated with or without 2 Gy IR. His-tagged Ub was transfected into PUMA KO PSCs, and then cells were lyzed for IP and IB analyses. c Effects of re-expression of PUMA in PUMA KO PSCs on Rad51 ubiquitination. PUMA cDNA (0, 2, or 4 μg) was transfected into PUMA KO PSCs. At 48 h after transfection, cells were treated with or without of 30 μM MG132 for 4 h. d Effects of PUMA knockout on Rad51 stability. At 8 h after IR, cells were treated with cycloheximide (CHX, 20 mg/ml) for the indicated time points. e Quantification of Rad51 protein levels in d. Data are representative of three independent experiments with similar results. Error bars, SD. **P < 0.01