Fig. 6

NF-κB binds to nos2 promoter to activate iNOS expression in myeloid cells. a The nos2 gene promoter structure. The NF-κB consensus sequence is indicated. The locations of ChIP PCR primers are also indicated. b ChIP analysis of NF-κB association with the nos2 gene promoter. c J774 cells were treated with LPS for approximately 1 h. Nuclear extracts were prepared from the cells and used for immunoprecipitation (IP) with anti-p65 and anti-p50 antibodies, respectively. The IP was then analyzed by Western blotting analysis using p65-specific antibody. d EMSA of NF-κB binding to nos2 promoter DNA. J774 cells were treated as indicated and used for nuclear extract preparation. The DNA probe containing the NF-κB consensus sequence as shown in A was incubated with the nuclear extracts and analyzed for NF-κB-DNA association using p65- and p50-specific antibodies