Figure 7
Effects of nitric oxide (NO) on Cyp19a1 expression in granulosa cells. (A) Expression of ovarian inducible nitric oxide synthase (iNOS) gene in DES-treated animals for the indicated times. Q-PCR data represent the mean ± SEM of at least four independent samples. Values marked by different letters are significantly different (P < 0.05). (B) Localization of iNOS and Cyp19a1 proteins in immature ovaries before (0 h) or after (6 h) DES injection. Both proteins were localized on granulosa cells. No staining was observed in control sections incubated with nonimmune serum. (C) Granulosa cells were isolated from immature DES-primed rats and treated with FSH (30 ng/ml) for the indicated times. Gene expression of each gene was measured by Q-PCR. Q-PCR data represent the mean ± SEM of at least four independent samples. Values marked by different letters are significantly different (P < 0.05). (D) Granulosa cells were isolated from immature DES-primed rats and treated with FSH (30 ng/ml) and DETA NONOate (50 μM) for 4 h. Gene expression of each gene was measured by RT-PCR. Uncropped images are shown in Supplementary Fig. S3. Effects of NO generator on expression of CYP19A1 mRNA (E) and proteins (F) in KGN cells. KGN cells were treated with 8-br-cAMP (1 mM) and DETA NONOate (50 μM) for 24 h. (E) CYP19A1 gene expression was measured by Q-PCR. Q-PCR data represent the mean ± SEM of at least four independent samples. Values marked by different letters are significantly different (P < 0.05). (F) Western blot analyses were performed with antibodies against CYP19A1 and GAPDH using lysates from KGN cells (30 μg protein) in each group. Uncropped blots are shown in Supplementary Fig. S3.
