Figure 4.

Effects of minodronate on growth and tube formation of microvascular EC. A, B, and E: ECs were incubated with or without 10 ng/ml of VEGF for 24 hours in the presence or absence of 100 nmol/L DPI, 1 mmol/L N-acetylcysteine, 10 μmol/L minodronate, 1 μmol/L GGTI-286, 1 μg/ml GGPP, or 10 μmol/L FTI-276, and then [³H]thymidine incorporation was measured. The percentage of [³H]thymidine incorporation is related to the value of the control. One hundred percent indicates 25,743 cpm. C and F: DN-RacT17N-, DN-RasS17N-, or mock-transfected ECs were treated with 10 ng/ml of VEGF for 24 hours, and then [³H]thymidine incorporation was measured. The percentage of [³H]thymidine incorporation is related to the value of mock-transfected cells without VEGF. One hundred percent indicates 5623 cpm. D: Ras activity. ECs were incubated with or without 10 ng/ml of VEGF for 24 hours in the presence or absence of 10 μmol/L minodronate or 100 nmol/L DPI. The percentage of band density is related to the value of the control. G: ECs were seeded on Matrigel and incubated with or without 10 ng/ml of VEGF for 6 hours in the presence or absence of 10 μmol/L minodronate. The percentage of length is related to the value of the control. One hundred percent indicates 4.65 mm/mm². * and **, P < 0.05 and P < 0.01 compared to the value with 10 ng/ml of VEGF alone, respectively (n = 4 to 6 per group). Similar results were obtained in two independent experiments.