Measuring ERCC1 and ERK1/2 Levels in Cancer Cells

In order to measure the levels of ERCC1 and ERK1/2, 5 × 10⁵ HROC277, HROC285, HROC370 or HROC374 cells per well were seeded in a 6-well plate, and treated by sham, 6.25 µM oxaliplatin, 5 mg/mL PRIVIGEN^® IgG, 5 mg/mL Tonglu^® IgG, 100 ng/mL IgG1 or the combination therapy indicated in the figure legends. After 36 h or 48 h, cells were harvested and western blot was performed as previously described using mouse anti-ERCC1 (Santa Cruz Biotechnology, Texas, USA, code sc-17809, dilution: 100×), mouse anti-ERK1/2 (R&D Systems, code MAB15761, dilution: 500×), rabbit anti-phospho-ERK1/2 (p-ERK1/2, R&D Systems, code MAB1018, dilution: 1000×) and mouse anti-β-actin antibody (Sigma-Aldrich, code A5441, dilution: 20,000×), followed by the secondary antibodies: peroxidase-linked anti-rabbit (Cell Signaling Technology, code 7074, dilution: 5000–10,000×) or anti-mouse antibodies (Sigma-Aldrich, code A9044, dilution: 20,000–60,000×). Proteins were visualised as previously described.^{29 (link)}