Depc treated h2o

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.

Recombinant human transforming growth factor-β1 (TGF-β1) was purchased from Peprotech (United States). Antibodies against collagen Ⅰ, fibronectin, P-Smad3/Smad3, P-Akt/Akt, P-ERK/ERK, P-JNK/JNK, P-P38/P38, GAPDH, E-cadherin, vimentin, and N-cadherin were purchased from Cell Signaling Technology (United States). The β-tubulin antibody was obtained from Proteintech (China). The α-SMA antibody was obtained from Santa Cruz Biotechnology (China). Goat pAbs against Rb IgG (HRP) and Rb pAbs against Ms IgG were obtained from ImmunoWay (China). TRIzol reagent was acquired from Ambion Life Technology (China). DEPC-treated H₂O and RNase Away H₂O were purchased from Life Technologies. Fastking gDNA Dispelling RT SuperMix was purchased from Tiangen (Beijing, China). UNICON® qPCR SYBR Green Master Mix and liposomal transfection reagents were purchased from Yevsen (Shanghai, China). The Masson’s trichrome staining kit was obtained from Solarbio (China). Recombinant Mouse Platelet-derived Growth Factor-BB were purchased from MedChemExpress (China).

Recombinant human transforming growth factor‐β1 (TGF‐β1) was purchased from Peprotech. Antibodies against fibronectin, P‐mTOR/mTOR, P62, LC3I/II were purchased from Cell Signalling Technology. Antibodies against α‐SMA, P‐PI3K/PI3K, P‐Akt/Akt, GAPDH were obtained from Affinity Biosciences. Goat pAbs against Rb IgG (HRP) and Rb pAbs against Ms IgG were obtained from ImmunoWay (China). TRIzol reagent was acquired from Ambion Life Technology (China). DEPC‐treated H2O and RNAse Away H₂O were purchased from Life Technologies. Fastking gDNA Dispelling RT SuperMix was purchased from Tiangen. UNICON® qPCR SYBR Green Master Mix and liposomal transfection reagents were purchased from Yevsen. The Masson's trichrome staining kit was obtained from Solarbio (China).