Fig. 4

OTUD3 interacts with p53. a-b Co-immunoprecipitation of endogenous OTUD3 and p53 proteins in MCF7 and DU4475 cells. MCF7 and DU4475 cell extracts were immunoprecipitated with anti-p53 (a) or anti-OTUD3 (b) antibodies, and rabbit IgG served as a negative control. c Overview of the OTUD3 structure and diagrams of the D1 and D2 constructs. A summary of their binding status with p53 is shown on the right. d Extracts of MCF7 cells transfected with the indicated constructs were subjected to immunoprecipitation with an anti-Myc antibody, followed by western blot analysis with the indicated antibodies. e A direct interaction between GST-p53 and OTUD3 and its truncated mutant was detected by GST pulldown experiments. f Overview of the p53 structures. A summary of their binding status with OTUD3 is shown on the right. g A Myc antibody was used for IP, and IB was performed to detect the interaction between OTUD3 and p53 and their truncated mutants in the co-IP experiment. h GST pulldown experiments were performed to detect the direct interaction between GST-OTUD3 and p53 and its truncated mutant