Integrin β1

Oridonin was obtained from Aladdin (Shanghai Biochemical Technology Co., Ltd.). PF573228 (FAK inhibitor) and PD98059 (ERK1/2 inhibitor) were purchased from Selleck. Roswell Park Memorial Institute (RPMI) 1640 medium and foetal bovine serum (FBS) was obtained from Grand Island Biological Company. TRIzol reagent was obtained from Invitrogen. 3‐(4,5‐Dimethylthiazol‐2‐yl) ‐2,5‐diphenyltetrazolium bromide (MTT) and dimethyl sulphoxide (DMSO) were purchased from Sigma. Primary antibodies against the following proteins were used: E‐cadherin, vimentin, snail and slug, which were purchased from Abcam Co., Ltd.; FAK, p‐FAK^(Tyr397), ERK1/2 and p‐ERK1/2^{(Thr202/Tyr204)}, which were obtained from Cell Signaling Technology Co., Ltd.; integrin β1 was purchased from ImmunoWay Biotechnology Company; and β‐actin and GAPDH were purchased from Santa Cruz Biotechnology Co., Ltd. Secondary anti‐mouse and anti‐rabbit antibodies were obtained from Pierce. Opti‐MEM serum‐free medium and lipofectamine RNAiMAX were obtained from Thermo Fisher Scientific.

The total protein in the cells was extracted using RIPA lysis buffer, and the concentration of the protein sample was determined using a BCA kit (Beyotime Biotechnology). Each sample underwent a 10‐min denaturing process at 100°C. Samples were separated using a 10% SDS polyacrylamide gel (Beyotime Biotechnology) and transferred to PVDF membranes (Millipore). The membrane was first blocked with 5% nonfat milk TBST buffer for 2 h, and primary antibodies were applied and incubated overnight at 4°C, followed by the application of secondary antibodies for 1 h. Blots were visualized using BeyoECL Moon (Beyotime Biotechnology). The antibodies used in this study included Fas (1:2000; R&D System), FasL (1:1000; Bioss), COL‐I (1:1000; Immunoway), integrin β1 (1:1000; Immunoway), Fibronectin (1:1000; Immunoway), RUNX2 (1:1000; Immunoway), Osterix (1:1000; Affinity), ALP (1:2000; Santa Cruz Biotechnology), LC3B (1:1000; Abcam), Beclin (1:2000; Santa Cruz Biotechnology), HRP‐conjugated secondary antibodies against rabbit and mouse (1:10000; Biosharp), and HRP‐labelled donkey anti‐goat IgG (1:1000; Beyotime).