FIG. 9.

ICER shRNA abrogates GnRH pulse frequency-dependent effects on FSHβ transcription. (A) Representative Western blot analysis using LβT2 cell lysates transfected with empty vector (pcDNA3) or an ICER II expression vector with either an shCtrl construct containing a nonspecific scrambled sequence or an shICER construct containing a sequence directed against ICER. (B) LβT2 cells were transfected with −140/+15 rFSHβLuc (4 μg) and with 2 μg of shCtrl (left panel) or shICER (right panel) constructs, followed by perifusion and stimulation with low (∧__∧; every 120 min) or high (∧∧∧; every 30 min) GnRH pulse frequencies. Bar graphs show the fold stimulation (means ± the SEM) relative to unstimulated levels. Significant differences (P < 0.05), as measured by one-way ANOVA with a post hoc Tukey-Kramer multiple comparison test, are denoted by different letters.