FIGURE 5:

Inhibition of EphA2-dependent cell retraction by cAMP requires S897 phosphorylation. (A) Representative images of phalloidin-labeled, EphA2-knockdown PC3 cell populations infected with pLVX-IRES-Neo lentiviral vector (Vector) or lentivirus encoding WT EphA2 (WT) or the indicated EphA2 mutants. The cells, pretreated or not for 40 min with 20 μM forskolin, were stimulated for 12 min with 0.5 μg/ml control Fc or ephrin-A1 Fc. (B) Histogram showing average cell areas ± SE for the different conditions (160 cells/condition from an experiment in which 80 cells/well from two wells were measured). ****p < 0.0001 for the comparison of ephrin-A1 Fc–stimulated cells with the corresponding Fc-stimulated cells by one-way ANOVA followed by Sidak’s multiple comparisons test. (C) Immunoblot of PC3 cells transduced with empty lentiviral vector control and cells expressing the different EphA2 mutants to assess the levels of EphA2 expression and phosphorylation on S897 and S901.