Figure 6.
EphrinA5FLAG expression in PC12 cells induces an additional activation of Akt after NGF treatment. A, Time course experiment to compare the response to NGF of two PC12 clones stably transfected with ephrinA5FLAG or the empty vector (Neo). Total cell lysates (40 μg of protein) were analyzed by Western blot to check the status of phosphorylation of TrkA and downstream signaling molecules Akt, Erk1/2, and PLCγ (supplemental Fig. 5, available at www.jneurosci.org as supplemental material). B, Densitometric analysis of the level of phosphorylation of Akt, Erk1, and Erk2 from four independent experiments. Statistical analysis was performed by Student's t test, *p < 0.05. C, Effect of EphA7–Fc addition on the activation of signaling cascades by NGF. Pools of PC12 cells stably expressing ephrinA5FLAG were pretreated with 1 μg/ml of EphA7–Fc or left untreated before NGF (100 ng/ml) addition for the time periods indicated. Total cell lysates were analyzed by Western blot as described in A.