Inhibition of Oncogenic Signaling Pathways

AZD9291 and PF02341066 (crizotinib) were purchased from Active Biochemicals (Maplewood, NJ). AZD6244 (selumetinib) and PD0325901 were purchased from Selleckchem (Houston, TX, USA). GSK1120212 (trametinib) was purchased from LC Laboratories (Woburn, MA). All agents were dissolved in DMSO at a concentration of 10 mM and aliquots were stored at −80°C. Stock solutions were diluted to the appropriate concentrations with growth medium immediately before use. Mcl-1, p-Mcl-1 (S159/T163), p-Bim (S69), caspase-8, PARP, p-ERK1/2 (T202/Y204), and ERK1/2 antibodies were purchased from Cell Signaling Technology, Inc. (Beverly, MA). Caspase-3 antibody was purchased from Imgenex (San Diego, CA). Bcl-2 antibody was purchased from Santa Cruz Biotechnology, Inc (Santa Cruz, CA). Bax and GAPDH antibodies were purchased from Trevigen (Gaithersburg, MD). Bim antibody was purchased from EMD Millipore (Billerica, MA). Actinomycin D (Act D), cycloheximide (CHX), mouse monoclonal anti-actin and anti-tubulin antibodies were purchased from Sigma Chemical Co. (St. Louis, MO).