Figure 6. Reduced NEMO expression prevents phosphorylation of nuclear IKKα, recruitment of IKKα to IL-12 promoters, and phosphorylation of histone H3 following LPS stimulation.

(A) Nuclear proteins from LPS-activated MoDCs were analyzed by Western blotting using antibodies against IKKα, anti–phospho-IKKα, and USF2. (B and C) ChIP assays were performed on MoDCs stimulated with LPS. IKKα (B) and phospho-Ser10 histone H3 (C) were immunoprecipitated using specific antibodies, and the κB site of IL-12p35 and IL-12p40 promoters was amplified. GAPDH was used as a negative control, and the input lane corresponds to 10% of total isolated chromatin amplified with corresponding primers.