Anti rock1 antibody

Total protein was extracted from SRA01/04 cells after lysis on ice using RIPA lysis buffer (Beyotime Institute of Biotechnology) and quantified using a BCA kit (Beyotime Institute of Biotechnology). A total of 30 µg protein samples per well were then transferred to PDVF membranes after resolving using 10% SDS-PAGE gels. Subsequently, the membranes were washed, blocked with 5% skimmed milk for 1 h and then incubated with the following primary antibodies at 4˚C overnight: Anti-E-cadherin antibody (1:10,000; cat. no. ab40772; Abcam), anti-N-cadherin antibody (1:5,000; cat. no. ab76011; Abcam), anti-Vimentin antibody (1:1,000; cat. no. ab92547; Abcam), anti-α-smooth muscle act in (α-SMA) antibody (1:1,000; cat. no. ab265588; Abcam), anti-RhoA antibody (1:5,000; cat. no. ab187027; Abcam), anti-ROCK1 antibody (1:1,000; cat. no. ab92547; Abcam) or anti-ROCK2 antibody (1:1,000; cat. no. ab134181; Abcam). After washing with PBS, the membranes were incubated with the HRP-conjugated goat anti-rabbit IgG secondary antibody (1:2,000; cat. no. ab6721; Abcam) for 2 h at room temperature. All antibodies utilized in the present study were purchased from Abcam. GAPDH was used as the loading control. The protein blots were visualized using enhanced chemiluminescence (ECL) reagent and densitometry analysis of the bands was performed using ImageJ (National Institutes of Health).