Goat anti mouse igg secondary antibody conjugated to alexa fluor 488

DMEM, FBS, goat anti-mouse IgG secondary antibody conjugated to Alexa Fluor 488, Fura-2 AM and all RT-qPCR reagents were obtained from Invitrogen (Carlsbad, CA). Neurotensin, insulin and phalloidin-TRITC were obtained from Sigma Chemical (St. Louis, MO). The geranylgeranyl transferase I (GGTase I) inhibitor GGTI 298, PI 3-Kinase inhibitor A66 and the MEK inhibitor PD0325901 were from R&D Systems (Minneapolis, MN). The dual PI3K/mTOR inhibitor NPV-BEZ235 was purchasedfrom Selleck Chemicals (Houston, TX). Primary antibodies used were as follows: YAP (H-9, sc-271134 and 63.1, sc-101199, final dilution 1:200 for immunofluorescence), tubulin (sc-5274; final dilution 1:400), actin (sc-47778; final dilution 1:400) and GAPDH (sc-365062; final dilution 1:400) (Santa Cruz Biotechnology); phospho-YAP Ser127 (D9W2I, 13008; final dilution 1:1000), YAP (15028; final dilution 1:1000 for western blots), phospho-p70 S6 KinaseThr389) (9205; final dilution 1:1000) and phospho-S6 Ribosomal Protein Ser240/244 (5364; final dilution 1:1000), phospho LATS Thr1079 (8654; final dilution 1:1000) and LATS2 (5888; final dilution 1:1000) were from Cell Signaling Technology (Danvers, MA). Horseradish peroxidase–conjugated anti-rabbit IgG and anti-mouse IgG were from GE Healthcare Bio-Sciences Corp (Piscataway, NJ). All other reagents were of the highest grade available.