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. 2007 Jun 28;104(28):11706–11711. doi: 10.1073/pnas.0700544104

Fig. 4.

Fig. 4.

DAK negatively regulates IFN-β signaling triggered by cytoplasmic poly(I:C) and SeV in 293 cells. (A) DAK inhibits cytoplasmic poly(I:C)-induced activation of ISRE and the IFNβ promoter. The 293 cells (1 × 105) were transfected with the indicated reporter (0.05 μg) and expression plasmids (0.5 μg of each). Sixteen hours after transfection, cells were further transfected with poly(I:C) (4 μg) (gray bars) or buffer (white bars) for 12 h before luciferase assays were performed. (B) DAK inhibits SeV-triggered activation of ISRE and the IFN-β promoter. The 293 cells (1 × 105) were transfected with the indicated reporter (0.05 μg) and expression plasmids (0.5 μg of each). Sixteen hours after transfection, cells were infected with SeV or left uninfected for 12 h before luciferase assays were performed. (C) DAK inhibits SeV-induced expression of IFN-β. The 293 cells (1 × 105) were transfected with empty control plasmid or an expression plasmid for DAK (0.5 μg of each). Twelve hours after transfection, cells were infected with SeV or left uninfected for 12 h before RT-PCR was performed with human IFNβ and β-actin primers. (D) Effect of DAK RNAi #4 plasmid on the expression of DAK. The 293 cells (1 × 106) were transfected with a control or DAK RNAi plasmid (5 μg) for 60 h before Western blot analysis was performed with the indicated antibodies. (E) DAK RNAi potentiates cytoplasmic poly(I:C)-induced activation of ISRE and the IFN-β promoter in 293 cells. The 293 cells (1 × 105) were transfected with the indicated reporter (0.05 μg) and DAK RNAi (0.5 μg) plasmid. Thirty-six hours after transfection, cells were further transfected with poly(I:C) (4 μg) (gray bars) or buffer (white bars) for 12 h before luciferase assays were performed. (F) DAK RNAi potentiates SeV-triggered activation of ISRE and the IFN-β promoter in 293 cells. The 293 cells were transfected as in A. The transfected cells were infected with SeV or left uninfected for 12 h before luciferase assays were performed. Relat. Lucif. Act., relative luciferase activity.

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