Anti her2

Cells were harvested and lysed in SDS lysis buffer (PMSF, Protease, and Phosphatase Inhibitor Cocktail added) for 30 min at 4°C. Total protein was extracted from tissues with T-PER Tissue Protein Extraction Reagent (Pierce, Rockford, IL, USA) according to the manufacturer's protocol. The proteins were dissociated and separated by SDS/PAGE and then transferred to PVDF membranes, which were incubated with primary antibodies. The primary antibodies used for western blotting and their sources were as follows: anti-SEMA3C (Proteintech, 19242-1-AP), anti-GAPDH (Proteintech, 60004-1-Ig), anti-p-ERK1/2 (Cell Signal Technology, #4370), anti-ERK1/2 (Cell Signal Technology, #4695), anti-p-EGFR (Cell Signal Technology, #3777), anti-EGFR (Proteintech, 18986-1-AP), anti-p-Her2 (Cell Signal Technology, #2244), anti-Her2 (Cell Signal Technology, #2244), anti-Her2 (Proteintech, 18299-1-AP), anti-p-MET (Cell Signal Technology, #3077), anti-MET (Proteintech, 25869-1-AP), anti-p-SRC (Cell Signal Technology, #6943), and anti-SRC (Proteintech, 11097-1-AP). Antigen-antibody complexes were detected using horseradish peroxidase-conjugated secondary antibodies (Cell Signaling Technology #7074; #7076) with enhanced chemiluminescence (ECL) western blot detection reagent (Merck Millipore) according to the manufacturer's protocol.

DHQ3 and 17-DR were obtained as described previously. They were dissolved in dimethyl sulfoxide (DMSO, Biosharp, Hefei, China) and stored at −20 °C. MG132, Nec-1, DAPI (4,6-diamidino-2-phenylindole), and 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) were purchased from Sigma-Aldrich (St. Louis, MO, USA). PI assay kits were purchased from Beyotime Institute of Biotechnology (Wuhan, China) and the Annexin V FITC/PI apoptosis detection kit was purchased from Nanjin KeyGen Biotech (Nanjing, China). The ATP Assay kit was purchased from Merck KGaA (Darmstadt, Germany). Lipofectamine 2000 was purchased from Invitrogen (USA). The following antibodies were used: anti-Mcl-1, anti-PARP, anti-RIP1, anti-RIP3 (Santa Cruz Biotechnology, Santa Cruz, CA, USA); anti-Bcl-2, anti-Bax, anti-HIF1a, anti-CDK4, anti-Her2, anti-EGFR (Proteintech, Chicago, IL, USA); anti-Hsp70, anti-Hsp90, anti-Akt (Cell Signaling Technology, Beverly, Massachusetts, USA); anti-caspase 3 and anti-caspase 8 (ENZO, Switzerland); anti-C-Raf (Abcam, Cambridge, MA, USA); and anti-β-actin (BioSharp, Hefei, China).