FIG. 2.

Binding of NF1 on a transient template does not require the binding of OTFs or the HREs. MMTV constructs were transfected into C127 cells for 24 h and nuclei were isolated. The nuclei were digested with DraI and ExoIII, and the purified DNA was then digested to completion by AlwNI. Ten micrograms of DNA was amplified by reiterative PCR using Taq polymerase and ³²P-end-labeled oligo-22. 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), mNF1, mOTF, mPHre, and mDHre LS mutant construct transfections. The 5′ boundary of NF1 is indicated.