Fig. 5.

Effect of eNOS overexpression on endothelial cell migration. PAEC were transiently transfected with either WT- or C94R/C99R-eNOS. After 48 h, eNOS protein levels (A) and basal NO generation were determined. A representative Western blot image is shown that verifies over-expression of both proteins. The blots were reprobed with HSP90 to confirm equal loading. Basal NO_x levels were also significantly increased in WT- and C94R/C99R-eNOS over-expressing cells (B). In the presence of H₂O₂ (400 µM, 1 h), NO_x levels were significantly attenuated in WT- but not C94R/C99R-eNOS over-expressing cells (B). H₂O₂ (400 µM) significantly attenuated wound closure in WT- but not C94R/C99R-eNOS over-expressing cells (C). Similarly, in HMEC-1 cells, cell migration was significantly increased in WT- and C94R/C99R-eNOS over-expressing cells under basal conditions (D). H₂O₂ (300 µM) significantly attenuated HMEC-1 cell migration in both WT- and C94R/C99R-eNOS over-expressing cells (D). However, migration was attenuated less in C94R/C99R-eNOS over-expressing cells (D). Data are mean±SEM, N=3–49, ^⁎p<0.05 vs. untreated; ^†p<0.05 vs. WT-eNOS.