Chemidoc touch imaging pc system

Cells were lysed with cell lysis buffer (Cell Signaling Technology, Inc., Danvers, MA, USA) containing protease inhibitor cocktail (Sigma-Aldrich). Total cell lysates were subjected to SDS polyacrylamide gel electrophoresis and transferred to polyvinylidene difluoride membranes by using an iBlot 2 Dry Blotting System (Thermo Fisher Scientific). Immunoblotting was carried out using the following antibodies; anti-caspase-3, anti-caspase-8, anti-cleaved caspase-8, anti-BIM, anti-Fas, and anti-β-actin antibodies (Cell Signaling Technology). Membranes were incubated with horseradish peroxidase-conjugated secondary antibodies (Cell Signaling Technology), and imaging was performed with a ChemiDoc Touch imaging PC system (Bio-Rad Laboratories, Inc., Hercules, CA, USA). All experiments were performed at least twice and the representative data of one experiment are shown.