Figure 1

TNF-α-induced activation and specific binding of NF-κB to the consensus andcd38 putative binding sites in HASM cells. Electrophoretic mobility gel shift demonstrating binding of nuclear proteins obtained from either control (untreated) or TNF-α-treated (50 ng/ml) HASM cells to labeled oligonucleotides corresponding the consensus (NF-κB-consensus) or putative cd38 (NF-CD38) NF-κB binding sequences. Note NF-κB binding (indicated by horizontal arrow) in samples obtained from TNF-α-treated cells. Binding specificity was confirmed using a 100-fold excess of unlabeled oligonucleotide corresponding to either the consensus or putative sequences. Binding to the consensus and putative cd38 sequences is abolished by excess unlabeled putative sequence (shown by vertical arrows). SP1 oligonucleotides were used as a nonspecific competitor to confirm the specificity of the binding. FP: Free Probe in this and subsequent figures; T: TNF-α. Representative of 4 different assays.