Effects of the different decoy ODNs on cytokine-stimulated CD40 expression.

(A) Statistical summary of the effects of the STAT-1, NF-κB, or IRF-1 consensus decoy ODNs (10 μM, 4 hours preincubation) on CD40 mRNA expression (calculated as percentage of the stimulated control) induced by TNF-α (100 U/mL) plus IFN-γ (1000 U/mL) in the cultured HUVECs (n = 5; *P < .05 versus TNF-α/IFN-γ). (B) Effect of the STAT-1 or IRF-1 consensus decoy ODN (10 μM, 4 hours preincubation) on TNF-α (100 U/mL) plus IFN-γ (1000 U/mL)–stimulated CD40 protein expression in HUVECs and THP-1 cells after 14 hours. Typical Western blot analyses are shown; qualitatively identical results were obtained with at least 2 further batches of endothelial or THP-1 cells. (C) Effects of the STAT-1, NF-κB, or IRF-1 consensus decoy ODNs on cell-surface expression of CD40 protein in HUVECs exposed to TNF-α (100 U/mL) plus IFN-γ (1000 U/mL) for 20 hours, as judged by flow cytometry. Data are expressed as mean fluorescence intensity (MFI). Representative histograms depict isotype control (dashed line), basal expression (solid line), and stimulated CD40 expression (solid line, area under the curve highlighted in gray). Qualitatively identical results were obtained with at least 3 further batches of cells.