Figure 3: Pattern of Akt phosphorylation in PC12 cells exposed to NGF in the presence or absence of NOS inhibitors. Cells were pretreated with NOS inhibitor 2 h prior to treatment with NGF. Representative immunoblots of lysates obtained from control and 20 mM L-NAME (a) or 2.0 mM S-MIU (c) in the presence or absence of NGF. Phosphorylated and nonphosphorylated forms of Akt are identified by the primary antibody indicated above each panel. Densitometric analysis of phosphorylated Akt normalized to total Akt levels in cells treated with NGF in the presence or absence of L-NAME (b) or S-MIU (d). NGF treatment significantly increased Akt phosphorylation above control levels in the presence or absence of NOS inhibitors (*, **, ***), and this increase was significantly higher in cells treated with NGF in combination with S-MIU as compared to those treated with NGF alone (+). This figure is representative of immunoblots obtained from 5 or 6 independent experiments.