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. 2012 Oct 16;287(50):41875–41887. doi: 10.1074/jbc.M112.421552

FIGURE 6.

FIGURE 6.

Glc-6-Pase plays a role in cAMP-induced steroidogenic gene expression. A, MA-10 cells were transfected with StAR-Luc (200 ng), 3β-HSD-Luc (200 ng), P450c17-Luc (200 ng), siGlc-6-Pase (100 pmol), and siRNA (100 pmol) for 24 h after transfection. Cells were serum-starved followed by cAMP treatment (100 μm). Experiments were performed in triplicate, and data are expressed in relative luciferase units (RLU) and as the fold activation relative to the control. Data represent mean ± S.D. of three individual experiments. B, MA-10 cells were transfected with StAR-Luc (200 ng), 3β-HSD-Luc (200 ng), and P450c17-Luc (200 ng) for 12 h after transfection. MA-10 cells were serum-starved and pretreated with S 3483 (1 and 10 μm) for 2 h, followed by treatment with cAMP (100 μm). Experiments were performed in triplicate, and data are expressed in relative luciferase units and as the fold activation relative to the control. Data represent mean ± S.D. of three individual experiments. C, MA-10 cells were transfected with siGlc-6-Pase (100 or 200 pmol) for 24 h after transfection, followed by treatment with cAMP (100 μm) for 12 h. Total RNA was isolated for semiquantitative RT-PCR analysis. Data represent mean ± S.D. of three individual experiments. D, total RNA was isolated from C samples for real time PCR analysis. Glc-6-Pase, StAR, P450scc and 3β-HSD were evaluated. Data represent mean ± S.D. of three individual experiments. E, MA-10 cells were pretreated with S 3483 (5 or 10 μm) for 2 h, followed by treatment with cAMP (100 μm) for 12 h. Total RNA was isolated for semiquantitative RT-PCR analysis. Data represent mean ± S.D. of three individual experiments.