Investigating Protein Signaling in ccRCC

Western blot assays were performed as described in our previous study [17 (link)]. Briefly, cultured cells or ccRCC tissues were lysed in RIPA lysis buffer, and protein concentrations were determined using the BCA assay kit according to the manufacturer’s protocols (Thermo Fisher Scientific, Waltham, MA, USA). The nuclear and cytoplasmic proteins from ccRCC cells were extracted using an NE-PER Nuclear and Cytoplasmic Extraction Kit (#78899; Thermo Scientific). The primary antibodies used were rabbit anti-RNF43 antibody (ab84125; Abcam), rabbit anti-LATS1 + LATS2 antibody (ab70565), rabbit anti-LATS1 + LATS2 (phospho T1079 + T1041) antibody (ab111344), rabbit anti-YAP antibody (ab52771), rabbit anti-YAP (phospho S127) antibody (ab76252), rabbit anti-GAPDH antibody (#5174), rabbit anti-β-actin antibody (#58169), and rabbit anti-histone H3 antibody (#4499), all of which were from Cell Signaling Technology (Danvers, MA, USA).