Figure 5. Itraconazole inhibits the AKT1-MTOR pathway. (A) U87 or C6 cells were treated with DMSO, indicated concentrations of itraconazole or 100 nM rapamycin for 36 h. Phosphorylation of MTOR (Ser2448), RPS6KB1 (Ser424 and Thr421), and EIF4EBP1 (Ser65 and Thr70) was examined by immunoblot. Total MTOR, RPS6KB, or EIF4EBP1 respectively was used as the internal control. The data are representative of 3 independent experiments. (B) U87 or C6 cells were treated with DMSO or the indicated concentrations of itraconazole. Phosphorylation of AKT1 (Ser473), CDKN1A (Thr145), and CDKN1B (Thr198) was examined by immunoblot. Total AKT1, CDKN1A, and CDKN1B, respectively was used as the internal control. The data are representative of 3 independent experiments. *P < 0.05; **P < 0.01; ***P < 0.001; Itra, itraconazole; Rapa, rapamycin.