Pretreatment of DCs with ACs inhibits DNA binding activity of NF-κB. NOD BMDCs (A) or sDCs (B) were pretreated with ACs at a ratio of 1:5 (DC/AC) for the indicated times or left untreated and then were stimulated with LPS (50 ng/mL) for 0.5 hours. Nuclear NF-κB DNA binding was measured via EMSA. Composition of NF-κB complexes was previously determined via supershift analysis. A double-stranded OCT-1 DNA probe was used as an internal control. Densitometric analyses represent the ratio of intensity of NF-κB to OCT-1 binding per unit area and are represented as arbitrary units for the respective EMSAs. (C) Nuclear NF-κB binding activity was measured in NOD BMDCs pretreated with ACs, polystyrene latex beads (LB), or necrotic cells (NC) for specified times or left untreated and then were stimulated with LPS for 0.5 hours. (D) NOD BMDCs were preincubated with cyclohexamide (10 μg/mL) (CH) for 15 minutes, treated with ACs for 3 hours, stimulated with LPS (50 ng/mL) for 0.5 hours, and nuclear NF-κB DNA binding activity was determined. For this and all other figures, the control lane AC represents data obtained with DCs plus ACs without stimulation (eg, LPS). Data are representative of 3 independent experiments.