FIGURE 4.

Wyeth inhibits the proliferative and tubulogenic activities of human endothelial cells. Human microvascular endothelial cells were: A, cultured in medium containing 2.5% fetal calf serum without Wyeth, or with Wyeth added at the indicated concentrations, and their proliferation was determined as described under “Experimental Procedures.” Values are averages ± S.D. of three experiments performed in quadruplicates. *, significant differences (p < 0.05) between untreated and Wyeth-treated cells. B, cultured in media containing 2.5% fetal calf serum (cnt) with or without Wyeth (25 μm) or a mixture of Wyeth (25 μm) and 11,12-EET or 14,15-EET (1 μm each), and their proliferation determined as above. C and D, plated onto Matrigel in SF medium (cnt) with or without Wyeth (25 μm) or a mixture of Wyeth (25 μm) and 11,12-EET or 14,15-EET (1 μm each). C, representative images of capillary-like structures taken 6 h after plating. D, capillary network formation was quantified as described under “Experimental Procedures,” and the values are averages ± S.D. of branches per microscope field calculated from three experiments each performed in triplicates. E, the levels of CYP2C9 mRNA in untreated and Wyeth-treated cells were analyzed by quantitative real-time PCR and normalized using the β-actin mRNA as reference. Values are averages ± S.D. calculated from six determinations done with reverse-transcribed products from two different experiments.