P fgfr3 y724

Human cancer cell lines DMS114 (small cell lung cancer; FGFR1 amplification) and RT112 (bladder cancer; FGFR3 fusion and amplification) were obtained from ATCC (Rockville, MD, USA) and CLS (Eppelheim, Germany), respectively during December 2012. All the cell lines (control and resistant) were authenticated at the University of Arizona Genetic Core facility (STR profiling) during February 2016. BGJ398 was purchased from Selleck Chemicals (Houston, TX, USA). Akt inhibitor, GSK2141795 was purchased from MedChem Express (Princeton, NJ, USA). Inhibitors were prepared as 10 mM stock solutions in dimethyl sulfoxide (DMSO). CellTiter-Glo reagent was purchased from Promega Corporation (Madison, WI, USA). Lipofectamine 2000 was obtained from Invitrogen (Carlsbad, CA, USA). Control and Akt siRNA were obtained from Cell Signaling Technology (Boston, MA, USA). Antibodies to phospho-FRS2α (y196), FGFR1, GSK3α, GSK3β, p-GSK3 α/β S9/S21, p-GSK3 β S9, p-Akt S473, p-Akt T308, pan-Akt, pMEK1/2, MEK1/2, pERK1/2, ERK1/2, pYAP-S127, YAP, TSC1, Cyclin B1, FOXM1 antibodies were obtained from Cell Signaling Technology. Antibodies against FGFR3, FRS2α, and GAPDH were obtained from Santa Cruz Biotechnology (Dallas, TX, USA). Antibodies to p-FGFR1 (y653/y654) from EMD Millipore (Billerica, MA, USA); and p-FGFR3 (y724) from Abcam (Cambridge, MA, USA) were also utilized.