Specificity of the decoy ODN technique in cultured HUVECs and THP-1 cells.

(A) Example of the specificity of the consensus decoy ODN for IRF-1. Cytokine-stimulated (100 U/mL TNF-α, 1000 U/mL IFN-γ) CD40 expression is significantly reduced in HUVECs pretreated with the consensus (IRF-1c) but not with the mutant (IRF-1m) decoy ODN. (B) Effects of the STAT-1 or NF-κB consensus decoy ODN on CD40 and E-selectin expression in HUVECs exposed to TNF-α (100 U/mL) plus IFN-γ (1000 U/mL) for 9 hours. Typical RT-PCR analyses are shown; comparable results were obtained in 2 further experiments each with different batches of cells. (C) Effect of a 4-hour preincubation with the IRF-1 consensus decoy ODN (10 μM) on nuclear translocation of IRF-1 in THP-1 cells exposed to TNF-α (100 U/mL) plus IFN-γ (1000 U/mL) for 3 hours. Typical EMSA is shown, with the appropriate supershift analysis (IRF-1 IgG) performed with a single batch of THP-1 cells. Identical results were obtained with another batch of cells.