Figure 6.

Pretreatment of confluent PAE cells with EGTA augments VEGF-induced tyrosine phosphorylation of VEGFR-2. Serum-starved confluent PAE cells overexpressing VEGFR-2 were treated with EGTA for the indicated periods and stimulated with VEGF for 5 min. Cells were lysed and immunoprecipitated with an anti-VEGFR-2 antibody. The immunoprecipitated proteins were collected, resolved on SDS-PAGE, and subjected to immunoblotting with an anti-phosphotyrosine (pY) antibody (A). The same membranes were reprobed with an anti-VEGFR-2 antibody (B).