FIGURE 7.

PI3K and eNOS signaling is involved in Fstl1-induced endothelial cell responses. A, effect of LY294002 on Fstl1-induced phosphorylation of eNOS and Akt. HUVECs were treated with LY294002 (10 μm) or vehicle following transduction with Ad-Fstl1 or Ad-β-gal. After 24 h serum deprivation, phosphorylation of eNOS (P-eNOS), and Akt (P-Akt) were determined by Western blot analysis. Representative blots are shown. Relative phosphorylation levels of eNOS and Akt were quantified (n = 4) by using ImageJ. Immunoblots were normalized to total loaded protein. B and C, contribution of PI3K to Fstl1-mediated endothelial differentiation and migration. HUVECs were treated with LY294002 (10 μm), l-NAME (1 mg/ml), or vehicle along with Ad-Fstl1 or Ad-β-gal for 8 h. After 24-h serum-starvation, Matrigel (B) or modified Boyden chamber assays (C) were performed. Results are shown as the mean ± S.E. (n = 5-8). Results are expressed relative to the values compared with control. ^*, p < 0.01.