Fibronectin 1

Total protein in the cells was extracted using a RIPA lysis buffer (Beyotime Institute of Biotechnology). After quantification using a bicinchoninic acid protein assay kit (Beyotime Institute of Biotechnology), protein (40 µg/lane) in the different experimental groups were separated via 10% SDS-PAGE, and then transferred onto PVDF membranes using electrophoresis. Following blocking with 5% skimmed milk for 1.5 h at room temperature, the PVDF membranes were subsequently incubated with primary antibodies overnight at 4°C. Next, the primary antibodies were removed and the membrane was incubated with the secondary antibodies for 1 h at room temperature. The primary antibodies against α smooth muscle actin (α-SMA; cat. no. ab7817; 1:1,000), collagen I (cat. no. ab34710; 1:1,000), collagen III (cat. no. ab184993; 1:1,000) and E-cadherin (cat. no. ab1416; 1:1,000) were obtained from Abcam, while the antibodies against SMAD3 (cat. no. 9523T; 1:1,000), fibronectin1 (cat. no. 26836S; 1:1,000) and β-actin (cat. no. 4970T; 1:1,000) were obtained from Cell Signaling Technology, Inc. The secondary antibodies (cat. nos. ab7090 and ab97040; 1:5,000) were purchased from Abcam.