Figure 2

Time-dependent VEGF stimulation of Src association with KDR/Flk-1. HUVECs were serum-starved and then treated with VEGF (15 ng/mL) for 0, 15, 30, 60, or 120 min. (A) Cell extracts from the indicated treatment conditions were immunoprecipitated using α-Src 2–17, and immunoblotted using α-KDR/Flk-1 (N-931) or α-Src 327. (B) Cell extracts from the indicated treatment conditions were immunoprecipitated using α-KDR/Flk-1 (N-931), and immunoblotted using α-phosphotyrosine (4G10), α-phosphotyrosine 419 Src, or α-KDR/Flk-1 (A-3). (C) Cell extracts from the indicated treatment conditions were immunoprecipitated using α-Src 2–17, and immunoblotted using α-Flt-1 (H-225) or α-Src 327. (D) Cell extracts from the indicated treatment conditions were immunoprecipitated using α-Flt-1 (H-225), and immunoblotted using α-phosphotyrosine (4G10), α-Src 327, or α-Flt-1 (H-225).