Figure 9.

FOXO1 suppression of GnRH-induced Fshb transcription does not involve regulation of c-Fos or c-Jun mRNA or protein levels. LβT2 cells were transduced with Ad-GFP or Ad-FOXO1-CA for 6 hours, then switched to serum-free media. Twenty-four hours after adenoviral infection, cells were treated with 0.1% BSA vehicle (Veh) or 10 nM GnRH for 1 hour, as indicated. Cells were harvested for total mRNA (A–D) or protein (E). (A–D) The results represent the mean ± SEM of three experiments performed in triplicate and are presented as amount of c-Fos mRNA relative to Gapdh (A), fold GnRH induction of c-Fos relative to Veh (B), c-Jun mRNA relative to Gapdh (C), fold GnRH induction of c-Jun relative to Veh (D). *, transcription is significantly repressed by FOXO1-CA compared to GFP using Student's t test. (E) Western blot analysis was performed on whole cell extracts using c-FOS, c-JUN, and FOXO1 primary antibodies and a horseradish peroxidase–linked secondary antibody. A representative image is shown.