Supplementary material for Teixeira et al. (1999) Proc. Natl. Acad. Sci. USA 96 (24), 13831-13838.
Fig. 8.
The distal SF-1 site of the rat MIS type II promoter is bound by SF-1 more avidly than is the proximal SF-1-binding site. Gel-shift experiments were done with radiolabeled duplex oligonucleotides corresponding to the distal (SF1-250, A and B) or proximal (SF1-200, C and D) SF-1 sites, excess competitor oligonucleotides, and R2C nuclear extract. (A) The distal site is incubated with no nuclear extract (Free), with 1 mg of nuclear extract (NE), and with 1 mg of nuclear extract with 5-, 10-, 25-, and 50-fold molar excess of distal SF-1 (SF1-250) (Left) or with 5-, 10-, 25-, and 50-fold molar excess of proximal SF-1 (SF1-200) (Right). (B) The gel-shifted bands were quantitated by a PhosphorImager and the volumes were plotted against competitor concentration. Competition with the distal site (closed circles) and the proximal site (open circles) is shown. (C and D) The experiments described in A and B were repeated with the proximal SF1 site (SF1-200) as a probe.