Figure 6.
EphA7-S inhibits B-cell migration and ephrin ligand-induced EphA7 phosphorylation. (a) Nalm-6 B-cells (2×105 cells/well) were migrated for 2 h through a 5 μM pore membrane to recombinant SDF-1α (100 ng/ml), with or without pre-incubation with recombinant soluble EphA7-Fc (EphA7-S). Data is expressed as percentage of cells migrating to SDF-1α alone (typically 15–20% of input cells). All conditions were performed in triplicate. One of four representative experiments is shown. (b) EphrinA4-induced tyrosine phosphorylation of EphA7-FL expressing Nalm-6 cells was measured either in the presence of soluble EphA7-Fc or Fc alone. Cells were stimulated for indicated time, after which whole-cell lysates were immunoprecipitated with anti-EphA7 antibody followed by Western blot with anti-phosphotyrosine antibody, then re-probed with anti-EphA7 to verify equal total EphA7-FL in each well.