Figure 3. Effect of Src and PI3K inhibition on PDGF-stimulated MAP Kinases in HLEB3 cells

Serum-deprived cells with and without inhibitor preloading were stimulated with PDGF (1 ng/ml) for 0, 10, 20, and 30 min, harvested and then lysed in lysis buffer. Cell lysates were applied on 10% SDS-PAGE, transblotted and probed with specific antibodies to P-ERK1/2, P-JNK, P-Akt, P-p38, and G3PD, respectively. A: Cells were stimulated with PDGF with and without preloading (30 min) of Src inhibitor, PP1, at 10 μM or 20 μM. B: Cells were stimulated with PDGF with or without preloading (30 min) of PI3K inhibitor, LY294002 at 15 μM or 40 μM. Cells without treatment of inhibitor but with stimulation of PDGF (1 ng/ml) were used as the control. The bar graph with averaged pixel values of the band intensities for each western blot is shown. Data presented are a typical representation of 2-3 separate experiments.