Fig 1: PUMA interacts with cytoplasmic Rad51 and EMI1. a IP and IB of PUMA interaction with EMI1, FBH1, and UCHL3 in PSCs and HPCs with or without IR. b IP and IB of PUMA interaction with EMI1 in HEK293T cells. c In vitro GST pull-down analysis. Purified GST-PUMA or GST proteins were mixed with cell extracts from PSCs. d Representative images of colocalization of PUMA and EMI1 in PSCs at 8 h post-IR. DAPI was used to show nucleus. An anti-EMI1 antibody (JG35-83, #NBP2-76833, Novus Biologicals) was used. Scale bars, 5 µm. e IP and IB of EMI1 interaction with cytosolic PUMA and Rad51 in PSCs with or without IR. f Effects of EMI1 KO with a sgRNA on PUMA-mediated Rad51 ubiquitination in PSCs and HPCs after IR. g Representative images of ?H2AX foci in PUMA WT and KO PSCs or HPCs transduced with or without an EMI1 sgRNA after IR. Scale bars, 10 µm. h Quantification of ?H2AX foci in g. The percentage of positive cells (=5 ?H2AX foci) was shown. Data are representative of three independent experiments with similar results. Error bars, SD. *P < 0.05, ***P < 0.001
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