Fig. 5 | Signal Transduction and Targeted Therapy

Fig. 5

From: PUMA facilitates EMI1-promoted cytoplasmic Rad51 ubiquitination and inhibits DNA repair in stem and progenitor cells

Fig. 5

PUMA promotes Rad51 ubiquitination by EMI1. a Schematics of PUMA WT and truncated constructs. b IP and IB of the interaction of PUMA mutants with EMI1 in HEK293T cells. Flag-tagged PUMA WT or the truncation mutants with HA-EMI1 was transfected into HEK293T cells. c IP and IB of the association of PUMA mutants with Rad51 in HEK293T cells. d Effects of ectopic expression of the D3 or D4 mutant of PUMA on EMI1-mediated Rad51 ubiquitination. Flag-tagged PUMA WT, the truncation mutants, or an empty vector with His-Ub, HA-Rad51, and V5-EMI1 was transfected into HEK293T cells. e IP and IB of PUMA interaction with Rad51 F129A mutant. f PUMA increases EMI1 association with Rad51 WT, but not the F129A mutant. V5-EMI1 and HA-Rad51 WT or F129A mutant with or without PUMA were co-expressed in HEK293T cells. g Effect of Rad51 F129A mutation on PUMA interaction with EMI1. h PUMA promotes EMI1-mediated the ubiquitination of cytosolic Rad51 WT, but not the F129A mutant. Data are representative of three independent experiments with similar results. Error bars, SD. *P < 0.05, **P < 0.01

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