Insulin like growth factor 1 igf 1

Rabbit anti-pAkt S473, anti-Akt, anti-pGSK3Β S9, anti-GSK, anti-IGF1R and anti-pIGF1R Y1135 monoclonal antibodies were purchased from Cell Signaling (Danvers, MA, USA). Mouse anti-GAPDH monoclonal antibody was purchased from Ambion (Carlsbad, CA, USA). Alexa Fluor® 488 goat anti-mouse IgG was obtained from Molecular Probes (Eugene, Oregon, USA). Horseradish peroxidase (HRP)-conjugated anti-mouse and anti-rabbit IgG were acquired from Sigma (Saint Louis, MO, USA). Mouse anti-β-catenin was purchased from Thermo Fisher (Waltham, MA, USA). Insulin-like growth factor 1 (IGF-1) was obtained from Cell Signaling and reconstituted in 20 mM citrate, pH 3.0, to obtain a stock concentration of 100 µg/mL. The 2-(4-morpholinyl)-8-phenyl-1(4 H)-benzopyran-4-one LY294002 (PI3K inhibitor) was obtained from Cell Signaling and diluted in dimethyl sulfoxide (DMSO) to obtain a stock concentration of 3 mg/mL. Recombinant Wnt3a was purchased from R&D Systems (Minneapolis, MN, USA) and diluted in phosphate-buffered saline (PBS) + 0,1% bovine serum albumin (BSA) to obtain a stock concentration of 200 µg/mL and a work concentration of 50 ng/mL.