Fig. 5

RNF19A targeted ILK as a ubiquitination substrate. (A) Cell lysates from T24 and 5637 cells were immunoprecipitated with IgG, anti-RNF19A, or anti-ILK antibodies, and the precipitates were then subjected to western blot analysis. (B) HEK-293T cells transfected with the indicated plasmids were lysed and then immunoprecipitated with anti-HA or anti-Flag antibodies. The precipitates were detected by western blotting. (C-D) qRT‒PCR was performed to detect ILK mRNA levels after RNF19A was overexpressed or knocked down. (E-H) western blot analyses of ILK protein levels in BCa cells with stable overexpression or knockdown of RNF19A and quantitative analyses. (I-L) The protein levels of ILK in T24 and 5637 cells with stable overexpression or knockdown of RNF19A after CHX treatment were measured via quantitative analyses. (M-N) An in vivo ubiquitination assay using ILK pull-down confirmed that the overexpression of RNF19A promoted the ubiquitination of ILK, whereas the knockdown of RNF19A had the opposite effect. *P < 0.05, **P < 0.01, and ***P < 0.001