Fig. 7.

SOX-C factors directly bind to each SOX-binding site of mouse GnRH intron A. EMSA was performed using in vitro translated SOX-C proteins. A, Probe sequences for EMSA. SOX-binding sites are indicated by boxes, and mutated nucleotides are shown as bold characters. B, In vitro translated SOX4 and SOX11 proteins (Myc-epitope-tagged forms) were detected by immunoblotting (IB) with an anti-Myc antibody (asterisks). Rabbit reticulocyte lysate (RRL) was used as a negative control. In vitro translated SOX4 (C) or SOX11 (D) proteins were incubated with ³²P-labeled wild type (WT) or various mutant probes as indicated at the bottom. Binding specificities of SOX-C factors to each probe were confirmed by the addition of 200-fold molar excess self-unlabeled probes as a cold competitor (Comp.).