Direct transcriptional regulation of <i>junb</i> by NF-κB in LPS-stimulated BMDCs.

<p>(A) <i>Structure of the</i> junb <i>gene</i>. <i>junb</i> has no intron and is approximately 1900 bp long. The 4 kB sites are located within a 200 bp domain located approximately 200 bp downstream of the <i>junb</i> polyadenylation signal. The grey bar with the inverted arrows (enh) indicates the enhancer-containing fragment which is amplified in ChIP experiments to visualize NF-κB binding. The black bar with the inverted arrows (cont) located downstream the NF-kB enhancer region indicates the amplified negative control fragment used in the ChIP experiments to exclude non-specific NF-κB binding. The numbers indicate nucleotide position with respect to the transcription initiation site (+1). (B) <i>NF-κB binding in LPS-stimulated BMDCs</i>. BMDCs were stimulated for various periods of time with LPS before ChIP analysis. NF-κB binding in the enhancer region is presented in arbitrary units as well as the parallel negative control ChIPs carried out with a preimmune serum (PI). Calculations were made with respect to the amplification of the “cont” negative control fragment. The presented data are representative of 3 independent experiments (see text). (C) <i>Inhibition of NF-kB binding at the junb enhancer in the presence of BAY11-7085</i>. The same experiment as in B was conducted except that BAY11-7085 or DMSO was added together with LPS to BMDCs and incubation allowed to proceed for 1 hour.