Western Blot Analysis of Apoptosis Markers

Western blot analysis was performed, as described previously (7 (link)) using the following: mouse anti-c-FLIP monoclonal antibody (NF6; cat.no ALX-804-428; Alexis Biochemicals, San Diego, CA, USA), mouse anti-caspase-8 monoclonal antibody (cat.no 9746; Cell Signaling Technology, Inc. Danvers, MA, USA), mouse anti-DR4 monoclonal antibody (cat.no ab13890; Abcam, Cambridge, UK), rabbit anti-DR5 polyclonal antibody (cat.no ab47179; Abcam), Pro-Apoptosis Bcl-2 Family Antibody Sampler kit (cat.no 9942; Cell Signaling Technology, Inc.) and Apoptosis Western Blot Cocktail (cat.no ab136812; Abcam). An enhanced chemiluminescence detection reagent (SuperSignal West Pico) was used for detection (Pierce Biotechnologies, Rockford, IL, USA) and α-tubulin and β-actin were used as loading controls. All western blots were representative of at least three independent experiments.