FIG 5.

CGP57380 and IGF1 treatments induce AKT(S473) phosphorylation but have the opposite effect on substrates without mTORC1. (A) HeLa cells treated with 10 μM CGP57380 (left) or 10 nM IGF1 (right) were harvested at the designated time points for immunoblotting. (B) Quantitation of p-AKT(S473)/AKT, p-S6K(T389)/S6K, and p-4EBP(S65)/4EBP ratios for CGP57380 (top) and IGF1 (bottom). (C) Lysates from cells expressing Dox-inducible MNK1(T334D) or MNK1(D191A) (Fig. 3B) were tested for phosphorylation of S6K(T389); the averages of 3 independent assays with SEM are represented for p-S6K(T389), corrected by dividing by total S6K values. The asterisks denote ANOVA-protected t tests (P < 0.05) comparing Dox induction to a no-Dox control.