Figure 1.

Aromatase mRNA, enzyme activity, and promoter usage in LSMCs treated with various aromatase inducers. Primary cultured LSMCs were starved in serum-free DMEM/F12 (1:1) medium for 12 h and treated with PGE₂ (100 nm), Bt₂cAMP (500 μm), Bt₂cAMP plus PDA (100 nm), DEX (250 nm), or DEX plus 10% FBS for 24 h. A, Aromatase mRNA expression levels were measured by real-time RT-PCR and were normalized to GAPDH mRNA. Levels are reported as fold change compared with cells containing vehicle only (vehicle) and represent the average result from LSMCs from eight subjects. B, Data shown are representative results from aromatase enzyme activity assays of LSMCs from one subject, performed in triplicate. The results from six subjects were reproducible, but the range of activity was different for each subject. All values are reported as the mean results of assays performed in triplicate, with the bars indicating the sem. *, P < 0.05 (ANOVA). C, CYP19 gene promoter usage with or without treatment in LSMCs was determined using multiplex RT-PCR. The relative usage of each promoter is reported as an average from six different subjects.