Figure 2: Temporal pattern for phosphorylation of Akt in NGF-treated IMR32 cells. (a) Representative immunoblots of protein lysates from control (untreated) and NGF- (50 ng/mL) treated PC12 cells separated by SDS-PAGE, transferred to nitrocellulose membranes. Phosphorylated and nonphosphorylated forms of Akt were detected by incubating membranes with phosphorylation-specific and non-phosphorylation-dependent antibodies, and the primary antibodies used are identified above each panel. (b) Densitometric analysis of phosphorylated Akt normalized to total Akt levels. NGF significantly increased Akt activation following 15 min of treatment (***). This figure is representative of immunoblots obtained from 6 independent experiments.