Figure 3
Effect of TNFα and IL-17 on AKT phosphorylation on Ser473. Cultures of HT-29 cells were incubated in serum-free medium for 24 h and then treated with vehicle, TNFα (25 ng/ml), IL-17 (50 ng/ml), or TNFα + IL-17. Western blots were prepared and probed with rabbit monoclonal antibody specific for phospho-AKT (Ser473) or whole AKT. A. Time course for AKT phosphorylation. In subsequent experiments (B - E) cells were treated for 15 min. B. Effect of EGFR inhibitor AG1478 (1 μM) or PI3K inhibitor LY294002 (20 μM) on AKT phosphorylation. C. Effect of Src inhibitor SU6656 on AKT phosphorylation. D. Effect of vehicle, TNFα, IL-17, or TNFα + IL-17 on AKT phosphorylation. E. Effect of vehicle, TNFα, IL-17, or TNFα + IL-17 on AKT phosphorylation, quantified by image analysis. Each bar represents the mean ± SE of four replicate cultures. Means with different letters were significantly different, P < 0.05.