USP45 interacts with Coro1B and mediates its ubiquitination. (A) Immunoprecipitation analysis of interaction between Flag-Coro1B and Myc-USP45 with Flag antibody. (B) Immunoprecipitation analysis of the interaction between Flag-Coro1B and endogenous USP45 by pulldown of Flag-Coro1B. (C) RT-PCR analysis of CORO1B mRNA level in control and USP45 knockdown HeLa cell. Quantification of CORO1B mRNA level was normalized to GAPDH. Data are shown as mean ± SEM from three independent experiments. (D) Immunoprecipitation analysis for Flag-Coro1B ubiquitination in HEK293T cells with expression of wild-type (WT) or mutant (C199A) Myc-USP45. Cells were lysed by a denaturing agent (1% SDS) containing buffer. Quantification of Ub was normalized to Flag-Coro1B. Data are shown as mean ± SEM from three independent experiments. Significance was determined using one-way ANOVA and Dunnett’s multiple comparisons test (C and D); ****P < 0.0001. Source data are available for this figure: SourceData FS3.