Figure 4.

Kinase-dependent and -independent role of EphA2 in ephrinA1-mediated inhibition of directional migration. A: Wound-healing migration assay. Confluent PC3 cells, expressing wild-type (WT) EphA2 or kinase-deficient mutants were serum-starved for 24 hours, scratched with a tip, and a photograph was taken (T₀). Complete medium was added to induce migration of cells in the wound, together with 1 μg/ml of ephrinA1-Fc or Fc alone, and after 24 hours photographs were taken. B: Boyden cell motility assay. PC3 cells expressing wild-type EphA2 or kinase-deficient mutants, after 24 hours of serum starvation, were seeded into the upper chamber of Boyden chambers. Cells were allowed to migrate for 24 hours toward the lower chamber filled with complete growth medium together with 1 μg/ml of ephrinA1-Fc. Cell migration was evaluated after crystal violet staining by counting cells in six randomly chosen fields. The results are representative of four experiments. *P < 0.001 stimulated versus control.