Fig. 6
From: USP15-dependent lysosomal pathway controls p53-R175H turnover in ovarian cancer cells
USP15 regulates p53-R175H levels in ovarian cancer cells. a, b Effect of USP7 knockdown on a p53-WT and b p53-R175H levels. c, d USP15 knockdown causes c no effect on p53-WT levels (ALST cells) but d decrease in p53-R175H levels (TYK-Nu and TOV-112D cells). e, f siRNA-mediated knockdown of USP15 has no effect on the levels of e p53-R273H in OVCA420 cells and f p53-R248Q in OVCAR3 cells. g USP15 co-immunoprecipitates with p53-R175H. h Increased ubiquitination of p53-R175H upon USP15 knockdown. p53-R175H was immunoprecipitated from TYK-Nu cells treated with non-targeting siRNA and USP15 siRNA, using anti-p53 antibody (FL-393). Immunoblot for ubiquitin (anti-Ub (P4D1)) was performed on the immunoprecipitated proteins. i USP15 knockdown by siRNA does not result in increased ubiquitination of p53-R273H in OVCA420 cells. j Lysosomal inhibitors reversed the effect of USP15 siRNA on p53-R175H levels. k MCB-613 treatment (6 µM, 2 h) results in decrease in USP15 protein levels
