Fig. 4.

Effects of acute endotoxemia on NF-κB nuclear binding activity. Neutrophils isolated from control (CTL) animals or 2 h after endotoxin (ETX) administration were treated with medium control (Med), IFN-γ, TPA, or TPA + IFN-γ (I + T) for 1 h. Nuclear extracts were analyzed for NF-κB binding activity using a gel retardation assay. Extracts prepared from medium treated cells isolated 2 h post endotoxin were incubated on ice for 15 min with antibodies to the p50 or p65 subunits of NF-κB (1 μg), or 40-fold excess of unlabeled cold competitor (Comp) prior to the labeled probe. One representative gel from three separate experiments is shown.