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. 2013 Jan 22;288(11):7467–7480. doi: 10.1074/jbc.M112.446401

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© 2013 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 2. — Activation of MEK1/2 and ERK1/2 in VEGF-stimulated endothelial cells is abrogated by pitstop. A, HUVECs were pretreated with vehicle or 10 μm pitstop for 15 min and then stimulated for the times indicated with 50 ng/ml VEGF. Cell lysates were analyzed by Western blotting for phosphorylated (Tyr(P)-1175) and total VEGFR2, phosphorylated (Tyr(P)-341) and total C-Raf, phosphorylated (Ser(P)-217/Ser(P)-221) and total MEK1/2, phosphorylated (Thr(P)-202/Tyr(P)-204) and total ERK1/2. B–E, quantification of phosphorylation is shown. White bars represent vehicle, gray bars represent pitstop. n = 3 independent experiments. *, p < 0.05; **, p < 0.01; ***, p < 0.001, ns = no significant difference. F, HUVECs were pretreated with the indicated concentrations of pitstop for 15 min and then stimulated with 50 ng/ml VEGF for 10 min. Lysates were analyzed by Western blot for phosphorylated (Thr(P)-202/Tyr(P)-204) and total ERK1/2.