Anti γ catenin

Western blot was conducted as previously described24 (link). Briefly, protein samples were prepared with RIPA lysis buffer (Sigma-Aldrich, Cambridge, MA, USA) with protease and phosphatase inhibitor cocktails (Thermo Fisher Scientific). After separation by sodium dodecyl sulfate polyacrylamide gel, proteins (30 μg) were transferred to a PVDF membrane (Millipore, Boston, MA, USA). After blocking with 5% nonfat milk, the PVDF membrane was incubated with primary antibody in a cold room overnight. After three washes with Tris-buffered saline Tween (TBST) for 15 min, the PVDF membrane was incubated with peroxidase-conjugated secondary antibody. Then after three washes with TBST for 15 min, the protein was visualized with ECL detection solution (Millipore). The primary antibodies were listed as follows: anti-CISD2, anti-GAPDH, anti-GSK3β, anti-p-GSK3β, anti-vimentin, and anti-N-cadherin (purchased from Sigma-Aldrich, St. Louis, MO, USA); anti-β-catenin, anti-p-β-catenin, anti-c-Myc, anti-E-cadherin, and anti-γ-catenin (purchased from Abcam, Cambridge, UK). GAPDH was used as internal control. The intensity of the protein bands was quantified with ImageJ and normalized to that of GAPDH.