FIG. 3.

Binding of NF1 to the MMTV promoter is required for the binding of OTF proteins. MMTV constructs (pMMTV-CAT and mNF1 mutant) were transfected into mouse C127 cells for 24 h, and the cells were treated with DEX for 1 h. Isolated nuclei were subjected to limited digestion with HpaII and ExoIII, and the purified DNA was digested to completion with HaeIII. Reiterative PCR was performed with 20 μg of these samples by using ³²P-labeled oligo-38. The PCR products were separated on an 8% denaturing polyacrylamide gel. (A) Schematic of the MMTV promoter as a transient template showing the restriction enzyme and PCR primer sites. (B) Samples from Wt (pMMTV-CAT) and mNF1 construct transfections. Lanes 5 and 10 are G sequencing tracks. The 3′ boundaries of NF1, OTF, and TBP are indicated.