Cortisol Secretion Pathway Figure 7. Effects of 8CPT-29-OMe-cAMP, Metabolites, and Sp-8CPT-29-OMe-cAMP on StAR mRNA Expression. AZF cells were incubated either without (control), or with 8CPT-29-OMe-cAMP, 8CPT-29-OMe-59AMP, Sp-8CPT-29-OMe-cAMP, or 8CPT-Ade at indicated times. Total RNA was isolated after indicated times. Membranes were hybridized with specific probe for bovine StAR mRNA as described in the Methods. Each lane contained 10 mg of total RNA. 18S rRNA bands from representative gels are shown as evidence of even loading. A) Concentration-dependent effect of 8CPT-29-OMe-cAMP on StAR mRNA. AZF cells were incubated without (control, white bar) or with 10–50 mM 8CPT-29-OMe-cAMP (grey bars) for 48 hr before isolating total RNA. B) Time-dependent effects of 8CPT-29-OMe-cAMP on StAR mRNA. AZF cells were incubated without (control, white bar), or with 8CPT-29-OMe-cAMP (50 mM, grey bars) for indicated times after which total RNA was isolated. mRNA is expressed as % control value at 72 h. C) Comparison of effects of 8CPT-29-OMe-cAMP, 8CPT-29-OMe-59AMP, and hydrolysis-resistant Sp-8CPT-29-OMe-cAMP on StAR mRNA. AZF cells were incubated without (control, white bar) or with either 8CPT-29-OMe-cAMP (30 mM, light grey bar), 8CPT-29-OMe-59AMP (30 mM, dark grey bar), or Sp- 8CPT-29-OMe-cAMP (30 mM, striped/grey bar) for 48 h after which total RNA was isolated. D) Concentration-dependent effect of 8CPT-Ade on StAR mRNA. AZF cells were incubated without (control, white bar) or with 10–50 mM 8CPT-Ade (grey bars) for 48 hr before isolating total RNA. doi:10.1371/journal.pone.0006088.g007 PLoS ONE | www.plosone.org 9 June 2009 | Volume 4 | Issue 6 | e6088