P egfr tyr845

LL-37 peptide was synthesized by Synpeptide Inc (Nanjing, China). We purchased 1,25(OH)₂D₃, KO-974 and Neratinib from MedChemExpress (NJ, USA). Protein A/G and primary antibodies against hCAP18/LL-37, ERK1/2, p-ERK1/2, MMP14, MMP9, N-cadherin, E-cadherin, Vimentin, Slug, VDR, EGFR, p-EGFR (Tyr845) and Snail were obtained from Santa Cruz (CA, USA). Antibodies against β-actin and p-HER2 (Tyr1248) were purchased from Abclonal (Wuhan, China), as well as anti-rabbit and anti-mouse secondary antibodies. Anti-flag was purchased from Proteintech (Wuhan, China). HER2 antibody was purchased from Cell Signaling Technology (MA, USA). The human HCC cell lines Huh7 and PLC/PRF-5 were purchased from American Tissue Culture Collection (ATCC, Shanghai, China). Cells were maintained in DMEM (Thermo Fisher Scientific, USA) supplemented with 10% fetal bovine serum (FBS), penicillin (100 U/mL) and streptomycin (100 mg/mL) (Solarbio, Beijing, China) at 37°C in 5% CO₂. We acquired 3,3’-diaminobenzidine (DAB) from Beyotime (Nanjing, China). All other reagents were analytical grade reagents and produced in China.

Growth factors (HGF and EGF) and tyrosine kinase inhibitors (gefitinib and erlotinib) were purchased from PeproTech Inc. (Rocky Hill, CT, USA) and Wako Pure Chemical Industries Ltd. (Osaka, Japan), respectively. A6730, 5-FU, SU11274, U0126, and Stattic were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA). Antibodies were obtained from the following sources and used at the indicated dilutions: Sigma-Aldrich Co. (St. Louis, MO, USA), UPP1 (1:600), p-STAT3, and α-tubulin (1:10,000); Cell Signaling Technology, Inc. (Danvers, MA, USA), p-c-Met Tyr1003, Tyr1234/1235, Tyr1349, p-EGFR Tyr845, Tyr1068, Tyr1173, p-Akt1/2, and p-Jak2 (all 1:10,000); Santa Cruz Biotechnology (Dallas, TX, USA), p-Erk1/2 (1:2000). Other reagents and biochemicals were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA).