Figure 7.
NF-κB decoy oligonucleotides as well as the targeting of c-jun attenuated Cox-2 induction after low-dose NO. (A) RAW 264.7 macrophages (1 × 106 cells) were exposed to oligonucleotides (3 μM) containing the murine Cox-2 promoter NF-κB site (decoy) or a mutated NF-κB site for 24 h. After the medium was changed, cells were stimulated with a combination of LPS/IFN-γ/NMMA (LPS, 10 μg/ml; IFN-γ, 100 U/ml; NMMA, 1 mM) or with GSNO (200 μM) for 15 h. Cox-2 expression was analyzed by Western Blot analysis. (B) Cells were transiently transfected and enriched as described in MATERIALS AND METHODS with an expression vector containing the sequence of a dominant-negative c-jun mutant to target c-jun (TAM-67 transf.). Cells were stimulated for 15 h with 200 μM GSNO or with a combination of LPS/IFN-γ/NMMA (LPS, 10 μg/ml; IFN-γ, 100 U/ml; NMMA, 1 mM). Cox-2 Western blot analysis was as described in MATERIALS AND METHODS. Blots are representative of three similar experiments.