Involvement of NF-κB and STAT-1 in caspase-11 expression. (A-B) CD11c⁺ DCs were treated with IFNβ (1000 IU/mL), or cytokine cocktail in the presence or absence of IFNβ (1000 IU/mL). At 15, 30, and 60 minutes, DCs were fixed, permeabilized, and analyzed by intracellular staining with (A) phospho-p65, total p65, (B) phospho-STAT-1, and total-STAT-1 antibody. (C) CD11c⁺ DCs were treated with LPS (1 μg/mL) for different time periods and analyzed for phospho-p65 and phospho–STAT-1 intracellular levels. (D-E) CD11c⁺ DCs were treated with IFNβ (1000 IU/mL), LPS (1 μg/mL), cytokine cocktail in the presence or absence of IFNβ (1000 IU/m) for 1.5 hours (D), or with LPS (1 μg/mL) for 4, 6, and 8 hours (E). Cells were fixed, sonicated, and subjected to ChIP analysis using antibodies to STAT-1, NF-κBp65, or control IgG. Precipitated DNA was isolated and evaluated by PCR using specific primers for the proximal regulatory region of the caspase-11 promoter. PCR products were fractionated through 2% agarose gel electrophoresis. Data are representative of 3 independent experiments (A-C) and 2 representative experiments (D-E).