Mmp2 and mmp9

Tissues or cells were lysed with RIPA buffer (Pierce, Rockford, IL) supplemented with protease inhibitors. Protein concentration was measured using a BCA protein assay (Thermo Scientific, Rockford, IL). Protein lysates, suspended in loading buffer, were separated on 10% SDS-polyacrylamide gels and transferred onto PVDF membranes (Millipore, Belford, MA). Then these membranes were blocked with 5% skim milk at room temperature for 1 h, and incubated with primary antibodies at 4 °C overnight. After washed, they were incubated with suitable HRP-conjugated secondary antibody at room temperature for 30 min and detected using an enhanced chemiluminescence (ECL) kit (Thermo Scientific). Antibodies for CASZ1, ERK and p-ERK were obtained from Abcam (Cambridge, MA), for RAF1, cyclinD1, E-cadherin, N-cadherin and Vimentin were purchased from Santa Cruz Biotechnology (Santa Cruz, CA), for MMP2 and MMP9 were purchased from Affinity Biosciences (Cincinnati, OH). β-actin antibody and corresponding secondary antibodies were purchased from Zhongshan Golden Bridge Biotechnology (ZSGB, Beijing, China).