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. 2011 Dec 1;6(12):e28454. doi: 10.1371/journal.pone.0028454

Figure 1. VEGF-induced activation of c-Src, Akt, but not VEGFR-2, PLCγ-1 and ERK1/2 requires NADPH oxidase-derived ROS.

Figure 1

(A) Protein extracts from HCAEC transfected with control siRNA (Scram-si) or si-p47phox were subject to Western blots as described in Materials and methods. Serum-starved HCAEC were treated with VEGF (50 ng/ml) for the times indicated. Membranes were sequentially blotted, stripped and re-probed with the phospho-specific antibodies as shown. Blots shown are representative of three independent experiments. (B) Same as in (A) except, the membranes were probed for phosphorylation of Y1175 VEGFR-2, Y783 PLCγ-1 and p42/44 ERK1/2. Anti-β-actin antibody was used as loading control.