Antibodies against erk1 2

Tricetin (purity ≥ 99%) was obtained from Extrasynthese, Genay, France. Recombinant rat IL-1β, MTT, and type II collagenase were purchased from Sigma-Aldrich, Merck KGaA, Darmstadt, Germany. Dulbecco's Modified Eagle's Medium (DMEM), penicillin, streptomycin, fetal calf serum (FBS), 0.25% pancreatic enzyme, TRIzol reagent, and cDNA synthesis kit were all purchased from Thermo Fisher Scientific Inc. PGE2 ELISA kit was purchased from R&D Systems, Minneapolis, MN, USA. Caspase-3 cellular activity assay kit, cell lysis buffer, and antibodies against ERK1/2, p-ERK1/2, p38, p-p38, JNK, and p-JNK were purchased from Cell Signaling Technology, Danvers, MA, USA. Antibodies against MMP-3, MMP-13, Bcl-2, and Bax were purchased from Abcam, Cambridge, USA. Antibody against MMP-1 was purchased from Proteintech Group Inc., Rosemont, USA. Griess reagent was purchased from Beyotime, Shanghai, China. SYBR Premix Ex Taq II was purchased from Takara Biotechnology, Dalian, China.

The monoclonal human CAP1 antibody was previously described50 (link). Antibodies against GAPDH and GSK3 beta antibodies were from Santa Cruz Biotechnology Inc. (Santa Cruz, CA). E-cadherin antibody was from BD Biosciences (Franklin Lakes, NJ). Snail antibody was from Abcam (Cambridge, MA), and antibodies against ERK1/2, phosphor-ERK1/2 (Thr202/Tyr204), phosphor-cofilin, phospho-GSK3α/β (Ser21/Ser9), and FAK and phosphor-Y397 FAK were from Cell Signaling Technology Inc. (Danvers, MA). Cofilin antibody was from the Cytoskeleton Inc. (Denver, CO). Cell lysates were prepared for Western blotting similarly as previously described6 (link)51 (link).

Hydrangenol was purchased from CoreSciences Co. (#BBP01679, purity 98.5%, Seoul, Korea). Human recombinant VEGF was obtained from R&D Systems (Minneapolis, MN, USA). Antibodies against ERK1/2, AKT, eNOS, VEGFR-2, phospho-ERK1/2, phospho-AKT, phospho-eNOS, and phospho-VEGFR-2 were purchased from Cell Signaling Technology Inc. (Danvers, MA, USA). Polyclonal antibodies against cyclin D1, cyclin E, CDK2, CDK4, p21WAF1, p27KIP1, p53, and GAPDH were purchased from Santa Cruz Biotechnology Inc. (Santa Cruz, CA, USA). Polyclonal antibody against MMP-2 was purchased from Chemicon (Temecula, CA, USA).

Western blotting was performed to evaluate the protein expression levels of DEGs. The liver tissues were homogenized using a liquid nitrogen pre-cooled high-speed tissue homogenizer (Gering Scientific Instruments Ltd, Beijing, China) and lysed using 10 μM phenylmethanesulfonyl fluoride (PMSF, Beyotime Institute of Biotechnology, Jiangsu, China) and a 1% protease inhibitor cocktail (104 mM AEBSF, 80 μM Aprotinin, 4 mM Bestatin, 1.4 mM E-64, 2 mM Leupeptin, and 1.5 mM Pepstatin A; Sigma). The protein contents were analyzed with the BCA Protein Assay Kit (Vazyme Biotech Co., Ltd, Nanjing, China). Standard Western blotting was performed, and the blots were visualized via a chemiluminescent system using ImageQuant LAS 500 imaging instruments (GE Healthcare Life Sciences, Shanghai, China) and quantified using the Image J analyzer software. Antibodies against ERK 1/2, p-ERK1/2, p-NF-kB p65, MAPK p38, p-MAPK p38, JNK, and p-JNK were purchased from Cell Signaling Technology (Danver, MA, USA). Antibodies against TLR4, NLRP3, IL-1b, MYD88, and NF-kB p65 were obtained from Abcam (Cambridge, MA, USA).