Hydroxyproline testing kit a030 2

BBD was obtained from Shanghai Pharmaceuticals Holding Co., Ltd, Shanghai, China, and dissolved in normal saline (NS). Diethylnitrosamine (DEN) was diluted in normal water at a concentration of 1:10,000 (v/v). Hydroxyproline Testing Kit (A030-2) was bought form Jiancheng, Nanjing, China. The antibody of α-smooth muscle actin (α-SMA) (ab5694, Abcam), Toll-like receptor 4 (TLR4) (ab22048, Abcam), pIKKα/β(2697, Cell Signaling), IKKα/β(sc-7607, Santa Cruz), pIκBα(2859, Cell Signaling), IκBα(4812, Cell Signaling), pERK1/2 (4370, Cell Signaling), ERK1/2 (4695, Cell Signaling), Cyclin D1 (ab134175,abcam) and GAPDH (AP0063, Bioworld) were used for western blot or cellular immunofluorescence staining. LPS (H178, Jiancheng, Nanjing, China), IL-6 (H007, Jiancheng, Nanjing, China), TGF-β1(H034, Jiancheng, Nanjing, China), TNF-α (H052, Jiancheng, Nanjing, China), MCP-1 (H115, Jiancheng, Nanjing, China) were used for endotoxin enzyme-linked immunosorbent assay (ELISA) test. LPS was purchased from Sigma.

BBD was obtained from Shanghai Pharmaceuticals Holding Co., Ltd, Shanghai, China. Hydroxyproline Testing Kit (A030-2) was bought form Jiancheng, Nanjing, China. (Dulbecco’s) modified Eagle’s medium (DMEM), fetal bovine serum (FBS), penicillin and streptomycin sulfate were purchased from Invitrogen (Carlsbad, CA, USA). The antibody of CK-19(10712-1-AP, proteintech), SOX-9(ab185230, Abcam), Toll-like receptor 4 (TLR4) (ab22048, Abcam), Ras (67648, CST), ERK (4695, CST), P-ERK (4370, CST) and GAPDH (AP0063, Bioworld) were used for western blotting assay or cellular immunofluorescence staining. The kit of endotoxin enzyme-linked immunosorbent assay (ELISA) test, including LPS (H178), IL-6 (H007), TGF-β1(H034), TNF-α (H052), MCP-1 (H115) were purchased from Jiancheng, Nanjing, China. LPS (L14130) was purchased from Sigma. Diethylinitrosamine (DEN) was obtained from Sigma-Aldrich, St. Louis, MO.