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Fig. 3. Time and voltage dependency of orientation of endothelial cells in a small physiological EF. (A) Orientation index as a function of time and voltage (n=65-598 from at least two independent experiments). (B) Effects of various drugs on EF-induced perpendicular orientation. Inhibition of PI3K (LY), Akt (Akt-i), Rho (Y27632) and actin polymerization (Latrunc) significantly decreased orientation responses, whereas inhibition of VEGFR (VEGFR-i), or combined inhibition of both Akt and Rho (Akt-i + Y) completely abolished the orientation response. VEGFR-i, VEGFR inhibitor (50 µM); LY, PI3K inhibitor LY294002 (50 µM); Akt-i, Akt inhibitor (50 µM); Y27632, Rho inhibitor (50 µM); Akt-i + Y27632, Akt and Rho inhibitors (10 µM each); Latrunc, Latrunculin (50 µM). Endothelial cells were subjected to EFs of 200 mV mm^–1 for 24 hours. n=47-343 from at least two independent experiments. **, P<0.0001 compared with cells exposed to 200 mV mm^–1 without drug treatment.

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