FIGURE 1.

Interaction of NF-κB (p50 and p65) with the GRK5 promoter region in cultured myocytes. A, putative consensus sequence for NF-κB binding with in 5′-flanking region of the GRK5 gene from different species. NF-κB binding site, GGGRNNYYC; R = A/G; N = any base; Y = T/C. B, competitive and antibody-mediated super shift EMSA using NE from NRVMs and labeled oligonucleotide for consensus NF-κB sites within the GRK5 promoter region. For EMSA super shift and competition assays, samples were incubated with specific antibodies to p50 or p65 and with 200-fold excess non-labeled consensus oligonucleotides, respectively, for 30 min prior to the addition of labeled nucleotides (see “Experimental Procedures”). Shown in lane 1 is the free probe; lane 2, NE alone; lane 3, NE + p50; lane 4, NE + p65; and lane 5, NE + 200× cold oligo (non-labeled). The experiments shown here is a representative of three independent experiments that yielded comparable results.