Figure 6. D-Asp-induced mTOR phosphorylation is a PI3K-PKB/Akt pathway-dependent event.

MGC were pre-incubated for 30 min with 10 nM of the PI3K inhibitor wortmannin (Wort) (A), with 100 nM of the PKB inhibitor (PKBI-IV) (B), or the p60^src inhibitor PP2 (10 nM) in the absence or presence of the RTK inhibitor genistein at 25 μM (C); and then treated with D-Asp (1 mM) or THA (100 μM). Levels of mTOR phosphorylation were detected as for Figure 1. (D) Levels of Tyr-527 phosphorylated p60^src were detected after treatment with 1 mM D-Asp in the presence/absence of W7 (25 μM) or genistein (25 μM). Phospho-p60^src levels were analysed by Western blot, Ponceau S was used as a loading control. The results are the mean ±S.E. of at least three independent experiments. In each panel, a representative Western blot is shown. Statistical analysis was performed comparing against non-stimulated cells using a non-parametric one-way ANOVA (Kruskal-Wallis test) and Dunn's post-hoc test (*P<0.05, ***P<0.001).