Figure 5.

Delayed DNA binding activity. EMSA showing that overexpression of mutant mE2* resulted in a delay in induction of NF-κB DNA binding activity in the HeLa cell nucleus. (A) HeLa cells were infected with control (empty) (lanes 1–4), wild-type mE2 (lanes 5–8), or dominant-negative mutant mE2* (lanes 9–12) expressing adenovirus vector, AxCA at a moi of 5 or 50. TNFα stimulation was done as in Fig. 4A. EMSA was performed with 5 μg of nuclear extract and ³²P-labeled NF-κB probe (lanes 1–12). Excess unlabeled wild-type κB competitor was added to the nuclear extract used in lane 13, and mutant κB unlabeled competitor was added to lane 14. (B) The intensity of NF-κB bands shown in A was measured by the PhosphoImager and indicated as bars. Lane numbers are corresponding to that of A. (C) EMSA with Oct1 was performed using the same nuclear extract used in A. Lane numbers correspond to those of A.