Characterization of NF-κB as a regulatory element in distal part of promoter.

<p><b>A)</b> Luciferase reporter gene assay for the effect of NF-κB and Twist-1 on the activity of <i>KIAA1199</i> promoter: Lysates of COS-1 cells transfected with cDNAs as indicated were examined by Dual Luciferase activity assay. Wild type and substitute mutation (pro-1.4 kb swap NF-κB^4th) at distal region of NF-κB binding site were used. <b>B)</b> ChIP assay for identification of the interaction between NF-κB (p65) and <i>KIAA1199</i> promoter. Top panel: A schematic diagram of four putative NF-κB binding sites relative to +1 site. Primer sets A, B, and C were designed for NF-κB binding site IV and I + II, and an area of the first <i>KIAA1199</i> intron, respectively. Middle and low panels: ChIP PCR in HeLa cells transfected with GFP control and P65 cDNAs. Anti-p-65 antibody and normal IgG (control) were used for immunoprecipitation. Results were calculated according to the bound/input ratio.