Fig. 5.

Determination of cAMP-responsive domains within the AMH promoter. A, Schematic representation of the human AMH gene-promoter-luciferase constructs. The longest construct contained 3068 bp. Localization and nucleotide positions of binding sites for NFκB, AP2, SF-1, AP1, SOX, and GATA-4 in the human promoter are according to Ref. 18 B, Cis-activating capacity of the constructs. KK1 cells were transfected with 1 μg of the previously mentioned constructs, and luciferase activity was analyzed after 48 h culture in control medium. Results are expressed in RLU. Means were compared by repeated-measures ANOVA, followed by Dunnett post hoc test to compare all vs. pGL2. C, Regulation of the constructs by cAMP. KK1 cells were transfected with 1 μg of each construct, and luciferase activity was assessed 48 h after treatment with cAMP (1 mm). Results are expressed as percentage of stimulation by cAMP compared with untreated cells. Means were compared by repeated-measures ANOVA and subsequently by Dunnett post hoc test to compare all vs. −3068hAMH-luc. Data shown correspond to the mean ± sem of at least three experiments, each done in triplicate. *, P < 0.05; **, P < 0.01.