Figure 7

The putative NF-κB and OCT1 binding sites in the upstream enhancing region are not required for promoter activity. A, EMSAs were performed with nuclear extracts and probes for the -1690/-1667 region of the promoter, resulting in specific complexes. Antibodies for p65, p50 and p52 subunits of NF-κB or rabbit IgG were incubated with the probe (arrow indicates supershift). Nuclear extracts were incubated with the -1737/-1707 probe and specificity of the complex was tested with unlabeled homologous, heterologous, consensus OCT and mutated consensus OCT oligonucleotides (arrow indicates specific complex). B, Cells were transfected with -5118pGL3, μNF-κBpGL3 or pGL3. In another experiment, cells were transfected with -5118pGL3, μOCT1pGL3 or pGL3. An asterisk indicates values that are greater than pGL3 (P < 0.05).