Formyl Peptide Receptor 1 Modulates Endothelial Cell Functions by NADPH Oxidase-Dependent VEGFR2 Transactivation

Figure 5

Tyrosine phosphorylation of 1175 residues of Flk-1/KDR triggers the PLC-γ1/PKC pathway. (a, c) Serum-deprived ECV304 cells were stimulated with 0.1 μM N-fMLP for the indicated times or (b) for 5 minutes in the presence or absence of PTX or apocynin. (a, b) Fifty micrograms of whole lysates was resolved on 10% SDS-PAGE, and PLCγ1 phosphorylation on the Y783 residue was detected by using an α-pPLCγ1 (Y783) antibody. An α-PLCγ1 antibody was used as a control for protein loading. (c) Membrane proteins (50 μg) were resolved on 10% SDS-PAGE, and PKC isoforms were detected by using the specific antibodies α-PKCα, α-PKCβII, α-PKCζ, or α-PKCδ as indicated. All the experiments are representative of at least three independent experiments. compared with unstimulated cells.

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