Monoclonal antibody to p53 and polyclonal antibodies to EGFR, MET, and AXL were from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Monoclonal antibodies to AKT1 and S6, and polyclonal antibodies to AKT, AKT2, and AKT3 were from Cell Signaling Technology (Beverly, MA, USA). Antibodies to MAPK, MDM2, and p21 were from Zymed/Invitrogen Laboratories (Invitrogen life Technologies, Carlsbad, CA, USA). All phospho-specific antibodies were from Cell Signaling Technology. Polybrene, puromycin, and antibody to β-actin were from Sigma-Aldrich (St Louis, MO, USA). Lentiviral shRNA constructs were from The RNAi Consortium (TRC, Cambridge, MA, USA), and included AXL: 5′-GCTGTGAAGACGATGAAGATT-3′; AKT1: 5′-CGCGTGACCATGAACGAGTTT-3′ (shRNA1), 5′-CGAGTTTGAGTACCTGAAGCT-3′ (shRNA2), 5′-CTATGGCGCTGAGATTGTGTC-3′ (shRNA3); and AKT2: 5′-CTTCGACTATCTCAAACTCCT-3′ (shRNA1), 5′-CAAGGTACTTCGATGATGAAT-3′ (shRNA2); and AKT3: 5′-AGAAACCTCAAGATGTGGATT-3′ (shRNA1), 5′-TGGCACACACTCTAACTGAAA-3′ (shRNA2).
Gefitinib, LY294002, U0126, GDC0941, and everolimus (RAD001) were obtained from LC Labs (Woburn, MA, USA). PHA-665752 was from Tocris Biosciences (St Louis, MO, USA). BEZ235 was from Axon MedchemBV (GZ Groningen, the Netherlands). All inhibitors were reconstituted in DMSO.
Gefitinib, LY294002, U0126, GDC0941, and everolimus (RAD001) were obtained from LC Labs (Woburn, MA, USA). PHA-665752 was from Tocris Biosciences (St Louis, MO, USA). BEZ235 was from Axon MedchemBV (GZ Groningen, the Netherlands). All inhibitors were reconstituted in DMSO.