A: patient-derived AML cell line that had been transfected with vector alone (TRI102), or the vector containing the tuberous sclerosis complex 2 (TSC2) gene (TRI103) were cultured with and without 10% FBS, and treated with or without RAD001 (20 nM, 24 h). Lysates were prepared, and tuberin, total- and phospho-S6 ribosomal proteins, and α-smooth muscle actin (α-SMA) were detected by Western blot. B: cell numbers were quantified under serum free conditions by crystal violet DNA dye binding in TRI102 and TRI103 cells treated with or without RAD001 (20 nM) for 24 h (n = 6), or 72 h (n = 10). C: cell death was measured under serum free conditions by propidium iodide uptake assay in TRI102 and TRI103 cells treated with or without RAD001 (20 nM) for 72 h or stuarosporine (ST; 250 nM) for 24 h (n = 3; **P < 0.01; ***P < 0.001). D: Western blot of apoptosis and cell cycle markers in TRI102 and TRI103 cells treated without (“C”) or with (“R”) RAD001 (20 nM) for 24 h. Results are represent 3 separate experiments. PARP, poly (ADP-ribose) polymerase; F.L., full length; Cl., cleaved.