Figure 8. IFN-αβ production after adenovirus induction is independent of known cytosolic nucleic acid sensors.

Wt and MDA-5^−/− BMDCs were infected with 0, 600, 1800 and 5400 Ad5GFP particles/cell, or stimulated with synthetic poly I∶C (10 µg/ml) or leader RNA complexed with Lipofectamine 2000 (0.1 µg/ml). IFN-αβ was measured in cell-free supernatants 16 h after infection (A). IRF-3^−/− BMDCs were transfected with pmaxGFP (1 µg/3×10⁵ cells) alone and together with RIG-IC (1 µg/3×10⁵ cells) or with DAI/Zbp1 targeting siRNAs (0.3 µM/3×10⁵ cells). Twentyfour hours later cells were infected with 5400 Ad5GFP particles/cell. Alternatively, RIG-IC transfected cells were stimulated with synthetic leader RNA (0.1 µg/ml) complexed with Lipofectamine 2000. Expression of IFN-β mRNA was measured in sorted GFP⁺ cells 12 h after stimulation. Knockdown of DAI/Zbp1 expression was measured in siRNA transfected cells (B).