Accumulation of u-PA, PAI-1, and t-PA in hormone-treated hMVEC-conditioned medium. hMVEC were cultured on three-dimensional fibrin matrices (for determination of u-PA and PAI-1 production) or gelatin-coated dishes (for determination of t-PA production) in incubation medium containing 20 ng/mL bFGF and 20 ng/mL TNF-α in the presence of vehicle (con) or the indicated hormones (1 μmol/L). (A and B) Show the accumulative production in the medium of u-PA (ng/well), (C and D) show the accumulative production of PAI-1 (ng/well), and (E and F) show the accumulative production of t-PA (ng/well). u-PA, PAI-1, and t-PA antigen were determined in the conditioned medium by ELISA as described in Materials and Methods. The data shown are from one representative experiment (of five performed). Under control conditions, the cumulative levels of u-PA, PAI-1, and t-PA after 9 days of culture varied between 20 to 49 ng/well, 2,300 to 2,440 ng/well, and 3.4 to 5.4 ng/well, respectively in the different experiments. For comparison, u-PA, PAI-1, and t-PA antigen levels were also determined when cells were cultured in incubation medium from which bFGF and TNF-α had been omitted (−BT). E2, 17β-estradiol; test, testosterone; prog, progesterone; dex, dexamethasone; 2-ME, 2-methoxyestradiol; D3, 1,25-dihydroxyvitamin D3; T3, thyroid hormone; at-RA, all-trans retinoic acid; 9-cis RA, 9-cisretinoic acid.