Mutation of FOXL2-binding sites reduces binding ability of FOXL2. A, Schematic of the −57-bp region of the CYP19 promoter illustrating the 30- and 57-bp wild-type (wt) and mutant (mt) oligonucleotide probes used. B, The 30-bp Wt1, Wt2, Mt1, or Mt2 probes were mixed with IP products from CHO cells transfected with the empty vector backbone (lanes 1–4), FLAG-tagged wild-type FOXL2 (lanes 5–8), or FLAG-tagged mutant FOXL2 (lanes 9–12). The protein-DNA complex band was detected in mixtures containing wild-type FOXL2 and either the Wt1 or Wt2 probes (lanes 5 and 6, asterisk-arrow) but not the Mt1 and M2 probes (lanes 7 and 8). This protein-DNA complex band was also strongly detected in mixtures containing mutant FOXL2 and the Wt1 or Wt2 probes (lanes 9 and 10) and more weakly detected in mixtures containing mutant FOXL2 and the Mt1 and Mt2 probes (lanes 11 and 12). C, The 57-bp probes Wt-57 bp, Mt1-57 bp, Mt2-57 bp, and Mt1/2-57 bp were also mixed with IP products from CHO cells transfected with the empty vector backbone (lanes 1–4), FLAG-tagged wild-type FOXL2 (lanes 5–8), or FLAG-tagged mutant FOXL2 (lanes 9–12). A protein-DNA complex band was strongly detected in mixtures containing wild-type FOXL2 and the Wt-57 bp probe (lane 5, asterisk-arrow) and more weakly detected in mixtures containing the Mt1-57 bp and Mt2-57 bp probes (lanes 6 and 7) but not in mixtures containing the Mt1/2-57 bp probe (lane 8). A protein-DNA complex band was strongly detected in mixtures containing mutant FOXL2 and the Wt-57 bp probe (lane 9, asterisk-arrow) and more weakly detected in mixtures containing mutant FOXL2 and the Mt1-57 bp, Mt2-57 bp, and Mt1/2-57 bp probes (lanes 10, 11, and 12, respectively).