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

Figure 3

Flk-1/KDR transactivation depends on NADPH oxidase activation. (a) Serum-starved ECV304 cells were stimulated with 0.1 μM N-fMLP for 5 minutes in the presence or absence of 100 μM apocynin, or (b) ECV304 cells serum-deprived for 24 hours were incubated for 12 hours with 5 nM siRNA against p22^phox (p22^phox siRNA) or negative control siRNA (NC siRNA) in DMEM containing 10% FBS in the presence of 20 μl of HiPerfect and stimulated with 0.1 μM N-fMLP for 5 minutes. Fifty micrograms of whole lysates were resolved on 10% SDS-PAGE and immunoblotted with α-pFlk-1 (Y951), α-pFlk-1 (Y996), α-pFlk-1 (Y1175), α-pFlk-1 (Y1214), or α-p22phox antibodies. An α-Flk-1 antibody served as a control for protein loading. All the experiments are representative of at least three independent experiments. compared with unstimulated cells. compared with unstimulated cells. compared with unstimulated cells. compared with unstimulated cells.

(a)

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