FIGURE 3. Effect of OXIM-11 on MIF regulation of glucocorticoid activity and on NF-κB activation from LPS-treated macrophages.

a, OXIM-11 decreased the level of TNF-α production in combination with dexamethasone (Dex) from MIF stimulation of LPS-treated human macrophages. Monocyte-derived macrophages from human peripheral blood were preincubated with dexamethasone (10⁻⁹ m) or dexamethasone plus MIF (100 ng/ml) and various concentrations of OXIM-11 before the addition of 0.5 μg/ml LPS. The data shown are mean ± S.D. of triplicate wells in experiments that were repeated twice. b, monocyte-derived macrophages from human peripheral blood were treated with various concentrations of OXIM-11 (1–100 μm) 30 min prior to LPS (10 ng/ml) addition. Nuclear extract was isolated using NE-PER nuclear and cytoplasmic extraction reagents. For detection of NF-κB binding, nuclear extract from cells (~5 μg of protein) was incubated with 0.2 ng of ³²P-labeled double-stranded oligonucleotide sequence, and the samples were resolved on a 4% polyacrylamide gel and visualized directly by autoradiography after drying the gel. The results for each treatment were expressed as relative intensity when compared with the control (saline-treated) intensity.