Comparable expression and phosphorylation of downstream targets of PI3K signaling. Cell lines were incubated with LY294002 or diluent alone (0.1% EtOH) for 15 minutes before lysis. Per sample, 40 μg protein was resolved by SDS-PAGE, transferred to nitrocellulose, and immunoblotted with antibodies to phosphorylated forms of Akt (A), FOXO (B), or GSK3β (C). Total Akt or total FOXO1 was used as a loading control. Pv indicates pervanadate; Exp, exposure to film: light (upper panel) and dark (lower panel). Data represent at least 3 different experiments and at least 2 different cell lines for each genotype.