Figure 4.

Effect of uPA on ezrin phosphorylation in the synapse. A and B, representative Western blotting analysis (A) and mean intensity of the band (B) of pERM, in synaptoneurosomes prepared from WT cerebral cortical neurons incubated for 60 min with 5 nm of uPA or a comparable volume of vehicle (control: C). The data are expressed as means ± S.E. for n = 4 observations. C–H, representative phosphate-affinity SDS–PAGE gel (C, E, and G) and mean intensity of the band (D, F, and H) of ezrin phosphorylated at Thr-567 and total ezrin (C, D, G, and H), and radixin phosphorylated at Thr-564 (E and F), in synaptoneurosomes (C and D) and whole-cell extracts (E–H) prepared from WT cerebral cortical neurons treated during 60 min with 5 nm of uPA or a comparable volume of vehicle (control: C). The data are expressed as means ± S.E. for n = 4 observations. I and J, representative Western blotting analysis (I) and quantification of the mean intensity of the band (J) of pERM expression in synaptoneurosomes prepared from WT neurons treated during 60 min with PBS or 5 nm of uPA, alone or in the presence of 10 μg/ml of anti–β3-integrin blocking antibodies. The data are expressed as means ± S.E. for n = 4–5 observations.