Fig. 5.

EMSA for Sp1/Sp3 binding to the CPI-17 promoter. A: scheme of mouse 169-bp DNA probe. B: EMSA for the biotinylated 169-bp probe (100 fmol) with recombinant Sp1 (rSp1, 100 ng) or the nuclear extracts from AoSMC (2 μg of proteins). Unlabeled DNA probe (2.5 pmol) was used as competitor. C: recombinant Sp1 binding to various DNA fragments, indicated in A. D: EMSA Supershift was performed using AoSMC nuclear extracts (2.4 μg) and the biotynylated 169-bp or 97-bp probe (100 fmol) in the presence of antibodies for Sp1 and Sp3 (1 μg). DNA-protein complexes are indicated at left. BLK, blank. Free probe (#) and supershift complex (*) with antibody are indicated.