FIG. 1.

NF-κB binding is inhibited in activated RAW 264.7 cells treated with 15dPGJ₂. (A) Macrophages were incubated for 1 h with different combinations of 15dPGJ₂ (2 μM), PGJ₂ (2 μM), rosiglitazone (10 μM), LPS (500 ng/ml), and IFN-γ (10 U/ml). After homogenization of the cells, nuclear extracts were prepared and the binding of nuclear proteins to the distal κB motif of the NOS-2 promoter was determined by EMSA. Supershift assays with Abs against proteins of the c-Rel family (not shown) identified p50-p50 and p50-p65 as the complexes present in the lower and upper bands, respectively. (B) Dose-dependent effect of 15dPGJ₂ on NF-κB activity. The binding of nuclear proteins to the PPARα R response element of the acyl-CoA oxidase promoter was used as control of extraction of nuclear proteins and lane load. The intensity of the bands was determined, and the corresponding values are expressed as mean ± SEM of three experiments (A). ∗, P < 0.005 with respect to the LPS/IFN-γ condition. a.u., arbitrary units.