Protein Expression Analysis in Liver Cancer

Tumor tissues and HCCLM3 and HepG2 cells were lysed using RIPA buffer (ThermoFisher Scientific, CA, USA) to obtain total proteins. Proteins were separated by SDS-PAGE and transferred to PVDF membranes. The above PVDF membranes were incubated with corresponding primary antibodies at 4°C overnight: anti-E-cadherin (1:1000; Cell Signaling Technology, MA, USA), anti-Vimentin (1:500; Sigma-Aldrich, MO, USA), anti-Tex10 (1:500; Proteintech Biotechnology, IL, USA), anti-Wnt1 (1:1000; Abcam, Cambridge, UK), anti-β-catenin (1:5000; Abcam), and anti-GAPDH (1:5000; Abcam). Then, the membranes were incubated with HRP-conjugated secondary antibody at 25°C for 2 h. Detection was conducted by enhanced chemiluminescence kit (ECL; ThermoFisher Scientific, CA, USA) [33 (link)].