Presence of estrogen receptors α and β, progesterone receptor, thyroid hormone receptors α and β, and vitamin D receptor mRNA in hMVEC as determined by RT-PCR. hMVEC were cultured for 4 days on gelatin-coated dishes in incubation medium containing 20 ng/mL bFGF and 20 ng/mL TNF-α or in incubation medium from which bFGF and TNF-α was omitted. RNA was isolated from these cells and cDNAs were synthesized using 1 μg total RNA and oligo dT primer as described in Materials and Methods. The cDNAs were amplified with primers for estrogen receptor α (A), estrogen receptor β (B), progesterone receptor (C), vitamin D receptor (D), and thyroid hormone receptors α and β (E) as described in Materials and Methods. The expected length of the amplified DNA fragment of the estrogen receptor α is 832 nt, of the estrogen receptor β 541 nt, of the progesterone receptor 737 nt, of the thyroid hormone receptor α 523 nt, of the thyroid hormone receptor β 458 nt, and of the vitamin D receptor 579 nt. As a positive control for the expression of estrogen receptor α, progesterone receptor, thyroid hormone receptors α and β, and vitamin D receptor mRNA, RNA isolated from MCF7 cells was used. As a positive control for the expression of the estrogen receptor β mRNA, RNA isolated from the SV-HFO osteoblast cell line was used. Lane 1, incubation medium from which bFGF and TNF-α had been omitted; lane 2, incubation medium containing bFGF and TNF-α; lane 3, appropiate positive control; M, molecular weight marker.