Figure 1.

Integrity of decoy ODN and AP-1 DNA binding. (A) Wild-type and mutated AP-1 decoy ODNs were denatured at 95°C for 5 min, cooled to room temperature for 2 h in the presence of annealing buffer, and then electrophoresed on a 20% polyacrylamide gel. DNA was visualized by ethidium bromide staining. Lanes 1, 4 and 5 are single-stranded ODNs; lanes 2 and 6 are double-stranded wild-type ODNs; and lanes 3 and 7 are double-stranded mutated ODNs. (B) AP-1-specific binding activity. AP-1 decoy ODN was biotin-labeled, 20 fmol was incubated in a binding reaction containing KB-3 cell nuclear extract and analyzed by gel electrophoresis, as described in Materials and Methods. The nuclear extract was omitted from the left lane, and the reaction in the right lane contained a 200-fold molar excess of unlabeled ODN.