Anti p15

Cells protein lysates were separated by 10% SDS-polyacrylamide gel electrophoresis (SDS-PAGE) transferred to 0.22μm NC membranes (Sigma) and incubated with specific antibodies. Autoradiograms were quantified by densitometry (Quantity One software; Bio-Rad). GAPDH antibody was used as control. anti-phospho-S6 kinase1, anti-phospho-S6, anti-p15 and anti-p16 were purchased from Santa Cruz Biotechnology, Inc. Anti-phospho-mTOR, anti-mTOR, anti-CDK6, anti-cyclinD1, anti-E2F1 and anti-Ki67 were purchased from Cell Signaling Technology, Inc.

Total-AKT, phospho-AKT (Thr308), total-ERK1/2, phospho ERK1/2, total PDK1, phospho-PDK1, Phospho-IGF-I Receptor β (Tyr1135/1136)/Insulin Receptor β (Tyr1150/1151) (19H7), total IGFR, PTEN and BRaf (55C6) were purchased from Cell Signaling Technology (Danvers, MA); Anti-p16INK4a antibody was purchased from Neo Markers (Fermont, CA); anti-mutated BRaf (V600E) antibody was purchased from New East Biosciences (Malvern, PA); anti-Grm1 antibody was purchased from R&D Systems (Minneapolis, MN); anti-p15 was from Santa Cruz; anti-β actin was from ThermoFisher; monoclonal -Tubulin antibody was obtained from Sigma (St. Louis, MO); anti- rabbit secondary was purchased from Merck ; anti-mouse secondary was purchased from Sigma (St. Louis, MO). Anti-sheep secondary was purchased from R&D Systems (Minneapolis, MN).

Cells protein lysates were separated by 15% SDS-polyacrylamide gel electrophoresis (SDS-PAGE) transferred to 0.22 μm NC membranes (Sigma) and incubated with specific antibodies. GAPDH antibody was used as control. Autoradiograms were quantified by densitometry (Quantity One software; Bio-Rad). Anti-p15 and anti-p16 were purchased from Santa Cruz Biotechnology, Inc. Anti-Ki67 was purchased from Santa Cruz Biotechnology.