Cell lysates were prepared in 1% NP-40 lysis buffer containing 1× cOmplete, Mini, EDTA-free Protease Inhibitor Cocktail (Roche, catalog no. 11836153001). Protein concentration was quantified using Pierce BCA Protein Assay Kit (Thermo Fisher Scientific, catalog no. 23225). Western blot samples were separated by SDS–polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to nitrocellulose membranes. The nitrocellulose membranes were blocked in 1× TBST (tris-buffered saline, 0.1% Tween 20) containing 5% BSA for 1 hour followed by incubation in primary antibody in blocking buffer overnight at 4°C.Blots were washed three times in 1× TBST for 5 min each, followed by incubation at room temperature for 1 hour in blocking solution containing appropriate secondary antibody (1:5000). Membranes were washed three times in 1× TBST and developed using Clarity Max Western ECL Substrate (Bio-Rad, catalog no. 1705062) and imaged by ChemiDoc MP Imaging System (Bio-Rad). GAPDH or E-cadherin was used as loading controls.

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