Figure 6.

Binding of EphA5–Fc to ephrin-A5-expressing cells induces changes in cell architecture, requiring the Src-family kinases. (A) NIH-3T3 cells expressing ephrin-A5 (A5-2) (a,b) and primary murine astrocytes (c,d) were incubated with Fc (a,c) or EphA5–Fc (b,d) proteins for 30 min at 37°C, fixed and subjected to indirect immunofluorescence using anti-vinculin monoclonal antibody (clone VIN-11-5, Sigma) and a Cy³-conjugated anti-mouse secondary antibody. (B) NIH-3T3 cells expressing ephrin-A5 (A5-2) (a–f) were incubated with Fc (a,c,e) or EphA5–Fc (b,d,f) proteins for 30 min at 37°C, in the presence of either vehicle (DMSO) (a,b), 1 μm Gö 6983 (c,d), or 10 μm PP1 (e,f). The cells were then fixed with methanol and subjected to indirect immunofluorescence using anti-vinculin monoclonal antibody.