Figure 4

The effect of PI-3-kinase and mTOR inhibitors on signal transduction pathways in HC11 cells. HC11 cells were serum starved overnight and incubated four hours with indicated inhibitors prior to re-stimulation with EGF (100 ng/ml) for times indicated. Lysates were harvested and analyzed by western blotting using antibodies specific for phosphorylated and non-phosphorylated forms of the indicated proteins. A. Cells treated with LY294002 (10 μM) and Rapamycin (50 nM) B. Cells treated with LY294002 (10 μM) and Rapamycin (50 nM) C. Cells treated with LY294002 (10 μM) and PD98059 (20 μM). D. The effect of conditionally activated-Akt1 on the Akt and p70S6K signal transduction pathway in HC11 cells. The HC11 cells were transiently transfected with either a conditionally active-Akt1 (CA-Akt) or a control vector. At 24 hours the cells were serum starved overnight in 4-hydroxy tamoxifen (1 μM). The next day the cells were re-stimulation with EGF (100 ng/ml) for times indicated. Lysates were harvested and analyzed by western blotting using antibodies specific for phosphorylated and non-phosphorylated forms of the indicated proteins.