Protein Expression Analysis by Western Blot

Protein expression was assessed by western blot analysis as we described previously (Xiao et al., 2015b (link)). In brief, protein (50 μg) samples were resolved by 10% SDS–PAGE and transferred to PVDF membranes, then probed with various primary antibodies against E-cadherin (1:500), α-SMA (1:500), COL-I (1:500) (Santa Cruz Biotechnology) and β-catenin (1:500) (Sigma-Aldrich, St Louis, MO, USA). After incubating with an appropriate secondary antibody, the protein expression was detected by western blot analysis, as described previously (He et al., 2009 (link)). β-actin (Santa Cruz Biotechnology, 1:1,000) was used as an internal control.