Fig. 2From: Pentoxifylline and Other Protein Kinase C Inhibitors Down-Regulate HIV-LTR NF-κB Induced Gene ExpressionSpecificity of the DNA-protein interaction involving a 32P-labeled oligonucleotide containing NF-κB motif and active NF-κB in a cell-free systemThe specificity of the interaction of active NF-κB with 32P-oligonucleotide in the PKC- or PKA-catalyzed cell-free activation system was determined by competition experiments with nonradioactive oligonucleotides carrying the wild-type (A) or mutated (B) NF-κB motif (described in Materials and Methods). The cell-free NF-κB activation system was as described in Fig. 1 with 1 µg of fractionated cytoplasmic protein, 20,000 cpm of 32P-labeled oligonucleotide, 0.2 U of PKC (designated by E) plus the indicated amounts (ng/5 µl reaction mixture) of nonradioactive wild-type or mutant oligonucleotides.Back to article page