Endomembrane H-Ras is mediating VEGF-induced migration by activating the PI3K/Akt/eNOS signaling cascade.A, in H-Ras-depleted ECs, reconstitution of endogenous H-Ras with either adenoviral H-Ras WT or endomembrane H-Ras PalM resulted in recovery of migration in response to VEGF. After overnight starvation, a wound healing assay was performed with H-Ras-depleted HAECs using VEGF (50 ng/ml) as a stimulus. Two-way ANOVA and Bonferroni's post-test were used (n ≥ 5). Error bars, S.E. ***, p < 0.001. B, reconstitution of endogenous H-Ras with either adenoviral H-Ras WT or H-Ras PalM resulted in partial recovery of the key proangiogenic signaling cascade Akt/eNOS in response to VEGF. HAECs treated with siH-Ras and reconstituted with H-Ras WT or PalM were stimulated after overnight starvation for 0 min, 10 min, 30 min, 4 h, and 24 h with VEGF (50 ng/ml). C, densitometric quantification of HAECs treated with siH-Ras and reconstituted with H-Ras WT or H-Ras PalM at 10-min VEGF stimulation. All values were standardized to GAPDH as a loading control and normalized to siC-LacZ at 0 min. n ≥ 3. Error bars, S.E.