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. 2018 Mar 18;2018:2609847. doi: 10.1155/2018/2609847

Figure 4.

Figure 4

FPR1-mediated Flk-1/KDR transactivation triggers the PI3K-Akt pathway. (a, c) Cell lysates were purified from serum-starved ECV304 cells exposed to 0.1 μM N-fMLP for the indicated times. (b, e) ECV304 cells were serum-deprived for 24 hours before the stimulation for 5 minutes with N-fMLP in the absence or presence of PTX, apocynin, (d) wortmannin, or LY294002. (f) ECV304 cells were serum-deprived for 24 hours, incubated for 12 hours with 5 nM siRNA against FPR1 (FPR1 siRNA) or (g) against p22phox (p22phox 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 was resolved on 10% SDS-PAGE and immunoblotted with (a, b) an α-pPI3K (p85) or (c–g) an α-pAkt (S473) antibody. An α-tubulin antibody served as a control for protein loading. All the experiments are representative of at least three independent experiments. ∗p < 0.05 compared with unstimulated cells. °p < 0.05 compared with unstimulated cells.