Figure 8.

FOXO1 suppression of basal Fshb transcription may involve FOXO1 regulation of Pitx1 mRNA and 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 harvested for total mRNA (A) or protein (B–C). (A) The results represent the mean ± SEM of seven experiments performed in triplicate and are presented as amount of Pitx1 mRNA relative to Gapdh. *, Pitx1 transcription is significantly repressed by FOXO1-CA compared to GFP using Student's t test. (B–C) Western blot analysis was performed on whole cell extracts using PITX1, FOXO1, and β-Tubulin primary antibodies and a horseradish peroxidase–linked secondary antibody. A representative image is shown (B) or graph representing the mean ± SEM of four experiments presented as the amount of PITX1 protein relative to β-Tubulin (C). *, PITX1 protein levels are significantly decreased by FOXO1-CA compared to GFP using Student's t test.