Figure 3.

Effect of NF-κB or GSK-3β inhibitors on binding of either c-Rel, RelB, or p50 subunits to a NF-κB consensus sequence. Heterophils were aliquoted into sterile 2-ml Eppendorf tubes (1 × 10⁷ cells/ml), where they were pre-incubated with the appropriate concentrations of the various inhibitors for 30 min at room temperature. Following these pre-incubations, the heterophils were then stimulated with S. Enteritidis (10⁹ cfu/ml) for 1 h at 41°C. The following inhibitors and optimal concentrations were used in these studies: BAY 11-7086 (IκB phosphorylation inhibitor; 50 μM), SN50 (NF-κB inhibitor, 100 μg/ml), and lithium chloride (LiCl, GSK3 inhibitor, 10 mM). Cell lysates (10 μg/ml) were used for binding of the activated c-Rel (A), RelB (B), and p50 (C) subunits to an NF-κB consensus sequence using the Trans-AM NF-κB ELISA kit. The experiment was performed in the presence of soluble wild type or mutated consensus oligonucleotides. The results are expressed as specific binding and are shown as mean ± SE of triplicate experiments. *p ≤ 0.01