Fig. 3.

Suppression of PGE2-induced the expression and promoter activity of aromatase by siRNA against ERRα. A, Cells in six-well plate were transfected with 25 pmol/well of siE or siC and grown for 24 and 48 h. ERRα mRNA expression was analyzed by real-time RT-PCR and protein levels were determined by Western blot. A, Upper panel, Real-time RT-PCR. Values represent mean ± sd (n = 3). **, P < 0.01 compared with untreated controls. A, Lower panel, Western blot analysis was performed using antibodies to ERRα or GAPDH. The experiments were performed in duplicate. Similar results were obtained from separate experiments. **, P < 0.01 compared with untreated controls. B, Cells in six-well plate were transfected with 25 pmol/well of siE or siC and then treated PGE2 (1 μm) or vehicle. After 24 h, total RNA was extracted and real-time RT-PCR was performed (B, upper panel). Values represent mean ± sd (n = 4). *, P < 0.05 compared with untreated controls. After 48 h, cells were lysated and Western blot was performed using antibodies to aromatase and GAPDH in duplicate (B, lower panel). Similar results were obtained from separate experiments. C, siE or siC were transfected into WPMY-1 cells together with the aromatase promoter luciferase reporter plasmid and then stimulated PGE2 (1 μm) or vehicle. Twenty-four hours later, luciferase activity was measured and normalized to renilla control activity. Results are the mean ± sd of three values. *, P < 0.05 compared with untreated controls.