Fig. 2.

Stimulation of the AMH promoter by cAMP, FSH, and LH in KK1 cells. Panel A, Stimulation of the −3068hAMH-luc construct by cAMP. Cells were transfected with 1 μg −3068hAMH-luc, and luciferase activity was assessed 24 and 48 h after treatment with cAMP (1 mm) and compared with cells cultured in control medium (C) using a Student's t test. Results are expressed in RLU or in percentage of stimulation [(RLU of treated cells − RLU of control cells)/RLU of treated cells) × 100]. Panel B, Effect of fetal calf serum (FCS) on the stimulation of −3068hAMH-luc by cAMP. Cells were transfected with 1 μg −3068hAMH-luc, and luciferase activity was assessed 48 h after treatment with cAMP (1 mm) in presence or absence of FCS and compared with cells cultured in control medium (C) using a Student's t test. Results are expressed in RLU. Panel C, Cis-activating capacity of −3068hAMH-luc in the presence of FSH-R and LH-R cDNAs. KK1 cells were cotransfected with 1 μg of the −3068hAMH-luc construct and 1μg of FSH-R and/or LH-R cDNAs, and luciferase activity was assessed after 48 h of culture in control medium. Results are expressed in RLU. Comparisons of means between different experimental conditions were made by repeated-measures ANOVA, followed by Dunnett post hoc test to compare all vs. pGL2. Panels D and E, Stimulation of the −3068hAMH-luc construct by FSH or LH. KK1 cells were cotransfected with 1 μg of the −3068hAMH-luc construct and FSH or LH receptor cDNA (1 μg). Luciferase activity was assessed after 48 h of treatment with FSH (panel D) or LH (panel E) (5–1000 mIU/ml). Results are expressed as percentage of stimulation. Responses to the different FSH and LH doses were compared by repeated-measures ANOVA and subsequently by Student-Newman-Keuls tests. Different superscripts indicate significant differences between the responses. Data shown correspond to the mean ± sem of at least three experiments, each done in triplicate. *, P < 0.05; **, P < 0.01; ***, P < 0.001.