FIG. 5.

EMSAs. (A) Double-stranded oligonucleotide sequences used in EMSAs. R3-a contains the R3-derived NF-κB sequence [termed NF-κB(R3)], and R3-b contains the mutant NF-κB sequence [termed NF-κB(TT)]. Putative PEA3 motif (AGGAAA) is conserved in either oligonucleotide. NFKB and NFKBm contain consensus or its mutant motifs for NF-κB, respectively. Each NF-κB motif is boxed, and mutated bases are indicated by open and solid triangles in R3-b and NFKBm, respectively. (B) Cold competition assays. The indicated molar excess of unlabeled competitor was added to the preincubation reaction mixture, and then ³²P-labeled probes were added. (C) Supershift analyses of NF-κB family bound to NFKB and R3-a probes with specific antibodies to p50, p65, and c-Rel. Nuclear extracts were preincubated with each antibody, and then ³²P-labeled probes were added. NFKBm was used to identify the specific complex.