Ripa lysis buffer middle

Cinobufagin (CBG) was purchased from Pusi Biotechnology Co. Ltd, and the purity of CBG was 99.21%. For each experiment, CBG was freshly prepared by dissolving in DMSO (Sigma, USA). Bleomycin (BLM) was acquired from Nippon Kayaku (Tokyo, Japan). TRIzol reagent was from Ambion Life Technology. DEPC TREATED H₂O, RNASE AWAY H₂O and SYBR Green Real-time PCR Master Mix were from Life Technologies. M-MLV Reverse Transcriptase was purchased from Promega, and PCR Buffer without MgCl₂ and MgCl₂ Stock Solution were acquired from Roche. In addition, Recombinant Ribonuclease Inhibitor, dATP, dTTP, dCTP, and dGTP were acquired from Takara. RIPA lysis buffer (middle) and the BCA kit were purchased from Beyotime Biotechnology. The primary antibodies described in the study include: anti-E-cadherin, anti-Vimentin, anti-fibronectin, and anti-collagen I (Affinity Biosciences, OH, USA) and anti-GAPDH, Smad3, P-Smad3, ERK, P-ERK, JNK, P-JNK, P38, P-P38 antibody (Cell Signaling Technology, Boston, United States); α-SMA and β-catenin antibody were from Santa Cruz Biotechnology (China) and Protein-Tech (China), respectively. The secondary antibodies anti-rabbit IgG (H+L) and anti-mouse IgG (H+L) were from Applygen (Beijing). Polyethylenimine, Linear purchased from Tianjin Hao Trading. β-catenin plasmid was purchased from MiaoLingPlasmid. ELISA kits were purchased from Beijing Suobao Technology.

Nintedanib (>99%) was purchased from HWRK Chem Co., Ltd. (Beijing, China). For each experiment, Nintedanib was freshly prepared by dissolving it in DMSO (Sigma-Aldrich, USA). Bleomycin (BLM) was acquired from Nippon Kayaku (Tokyo, Japan). Wnt3a was purchased from Peprotech (Texas, USA). TRIzol reagent and DEPC-treated water were obtained from Thermo Fisher Scientific corporation (Waltham, USA). RNase, DNase, and DNA Away H₂O were purchased from Beyotime Biotechnology (Beijing, China). FastKing gDNA Dispelling RT SuperMix was obtained from Tiangen Biotech Co., Ltd. (Beijing, China). The inhibitor KX2-391, RIPA lysis buffer (middle) and the BCA kit were purchased from Beyotime Biotechnology (Beijing, China). The primary antibodies described in the study included anti-fibronectin, anti-collagen I, and anti-β-tubulin antibodies (Affinity Biosciences, USA); anti-GAPDH, anti-p(Y416)-Src, and anti-Src antibodies (Cell Signalling Technology, USA); anti-α-actin antibody (Santa Cruz Biotechnology, USA); anti-β-catenin and anti-lamin B1 antibodies (ProteinTech Group, China); and anti-p(Y654)-β-catenin antibody (Immunoway Biotechnology, China). The secondary antibodies HRP-labeled goat anti-rabbit IgG and HRP-labeled goat anti-mouse IgG were from Abcam (Cambridge, UK).