Figure 7

Activin A stimulates, and TGFB1 inhibits, Fshb transcription in gonadotropes. Purified gonadotropes (A) or whole pituitary cells (B) were plated in triplicate for control, activin A, or TGFB1 treatments in 96 well culture plates. After 24 hrs, cells were pre-treated with follistatin (250 ng/ml). After 24 hrs, all media was removed, and the cells were treated with control media, or media containing either activin A (60 ng/ml) or TGFB1 (60 ng/ml). After 24 hrs of treatment, total RNA was isolated and real-time RT-PCR was performed. Normalized threshold cycle (Ct) values were averaged and used to quantitate Fshb mRNA expression with the 2^-(ΔΔCt) method for quantitation. The mRNA levels were quantitated relative to the mRNA levels of Fshb in control cells for purified gonadotropes (A) or whole pituitary cells (B), which were normalized to 100 %. Plotted are the means (+/- SEM) for three experimental replicates. Bars with different letters differed significantly.