Figure 6.

FOXO1 does not bind directly to the proximal Fshb promoter. TnT Flag-FOXO1-CA was incubated with a consensus FBE, −95/−61, −65/31, or −35/−1 Fshb probes and tested for complex formation in EMSA. (A) FOXO1-CA-DNA complex on the FBE is shown in lane 1, while anti-Flag supershift is shown in lane 3 and IgG control in lane 2. The FOXO1-CA-DNA complex (arrow), antibody supershift (ss), and nonspecific binding of proteins (ns) are indicated on the left of the gel. (B) Self-competition with excess cold FBE is shown in lane 14, lack of competition with mutant FBE, −95/−61, −65/31, or −35/−1 Fshb oligos in lanes 15–18.