Anti mmp2

A mouse anti-α-SMA (1:400) antibody was purchased from Boster (Wuhan, China). Rabbit anti- E-cadherin (1:400), anti-MMP2 (1:400), anti-MMP9 (1:400) were obtained from Boster (Wuhan, China). Rabbit anti-PTEN (1:500) were obtained from Bioss(Woburn, USA). Rabbit anti- pAKT (1:200), anti-AKT (1:200), anti-β-actin(1:1000) were obtained from Santa Cruz (Dallas, USA). Briefly, the cells were lysed on ice in a RIPA lysis buffer (Beyotime, China) and then denatured. The amount of protein was quantified by the BCA protein assay kit (Beyotime, China). SDS-PAGE was used to separate equal quantities protein samples and then proteins were transferred to polyvinylidene fluoride membranes. After blocking with 5% dried skimmed milk in PBS for 1 h, the membranes were incubated with various specific primary antibodies respectively, at 4 °C overnight. Then they were incubated with secondary antibodies. The amount of bound antibody was detected by ECL detection reagent(Beyotime, China). The relative optical densities of the bands were measured to compared the expression levels of the target proteins among different groups. β-actin was served as a loading control.