Western Blot Analysis of EMT Markers

The procedures for western blotting analysis and protein visualization were executed as described previously [24 (link)]. Primary antibodies against DVL3 and β-catenin were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Primary antibodies against E-cadherin, ZO-1, N-cadherin, Vimentin, Snail, CD44, CD133, SOX2, c-Myc were gained from Cell Signaling Technology (Danvers, MA, USA). Anti-β-actin (Santa Cruz, CA, USA) was used as a loading control. The horseradish peroxidase (HRP)-conjugated secondary antibody was obtained from ZSGB-bio (Peking, China). The signals were checked by an enhanced chemiluminescence detection kit(Invitrogen, Carlsbad, CA, USA) and Bio-Rad Molecular Imager (Hercules, CA, USA). The relative expression of above proteins was normalized against β-actin using Image J analysis.