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Fig. 4. Ena/VASP proteins are required for repulsion from substrate-bound ephrin stripes. (A-D) Wild-type MEFs, VASP^–/– MEFs and MV^D7 cells bind pre-clustered ephrin-A5-Fc, demonstrating Eph receptor expression (A,C,D); pre-clustered human Fc, applied as a negative control, was not bound by these cells (wild-type MEFs shown in B). (E) The loss of VASP and/or Mena does not prevent the activation of endogenous Eph receptors: EphA4 immunoprecipitations showed an increase in phospho-tyrosine (p-Y) following stimulation with 1 µg/ml pre-clustered ephrin-A5-Fc for 15 minutes, compared to unstimulated cells (0 minute) for each cell type. (F) The repulsive responses of VASP^–/– MEFs and MV^D7 cells were reduced compared to those of wild-type MEFs. Results are the means and s.e.m. from three or four experiments, with 600-800 cells counted for each condition. Means are significantly different from Wt MEF controls (denoted by ^*), according to Student's t-test [P<0.05, n=3 (VASP^–/– cells) or 4 (MV^D7 cells)]. Bars, 20 µm (A-D).

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