Anti mmp7

The cells were harvested and lysed using cell lysis buffer [with phenylmethylsulfonyl fluoride (PMSF)]. Then the lysates were repeatedly pumped with 1-ml injectors and sonicated with 3–4 bursts of 5–10 sec each. Finally, the protein lysates were mixed with β-mercaptoethanol and heated in a boiling water bath for 10 min. Next, the protein lysates were subjected to SDS-PAGE, transferred to PVDF membranes, and then blotted according to standard methods with anti-ACVR1 (1:800; cat. no. 4398; Cell Signaling Technology, Inc., Danvers, MA, USA), anti-p-β-catenin (Ser657) (1:500; cat. no. 4176; Cell Signaling Technology), anti-β-catenin (1:300; cat. no. 610153; BD Biosciences, San Diego, CA, USA), anti-cyclin D1 (1:500; cat. no. 60186; ProteinTech Group, Inc., Chicago, IL, USA), anti-MMP7 (1:200; cat. no. 10374; ProteinTech Group). Anti-α-tubulin monoclonal antibody (Sigma-Aldrich) served as a loading control. All the membranes were incubated with the HRP-conjugated secondary antibody (1:2,000; anti-mouse IgG; cat. no. 7076; 1:2,000; anti-rabbit IgG; cat. no. 7074; CST Shanghai Biological Reagent, Co., Ltd., Shanghai, China). At last, the protein bands were detected by enhanced electrochemiluminescence (Tanon Science and Technology, Co., Ltd., Shanghai, China) with SuperSignal West Pico (Thermo Fisher Scientific, Inc.).