Fig. 6

CRE decoy oligonucleotides decreased cAMP-mediated protection. (A) RAW 264.7 macrophages (1 × 10⁶ cells) were incubated with decoy or control olignucleotides for 24 hr as described under Materials and Methods. After changing the medium cells were prestimulated with 1 mM db-cAMP for 15 hr as indicated. Afterwards, DNA fragmentation was initiated by the addition of 1 mM GSNO or 0.25 mM Sp-NO. DNA fragmentation was quantitated after 8 hr by the diphenylamine assay as described in Materials and Methods. Data are mean values ± SD of three individual experiments (*p ≤ 0.05 versus control oligonucleotides). (B) Following prestimulation as described, p53 accumulation was determined after 4 hr by Western blot analysis as described in Materials and Methods after addition of NO donors. The blot is representative of three similar experiments.