FIG 7.

The MERS-CoV N protein interacts with TRIM25 and inhibits RIG-I interferon signaling. (A and B) 293T cells were transfected with 2 μg of the indicated plasmids for 36 h. Whole-cell lysates were subjected to anti-Flag immunoprecipitation and immunoblotting with anti-Flag or anti-GFP antibodies. (C and D) 293T cells were cotransfected with the IFN-β-Luc firefly luciferase reporter plasmid, the Renilla luciferase control reporter plasmid pRL, Flag-2CARD, and increasing amounts of the GFP-MERS-N plasmid (C) or GFP-MERS-N (aa 1 to 170) or GFP-MERS-N (aa 171 to 413) (D) for 36 h. The luciferase activity of the cell lysates was analyzed with the dual luciferase reporter assay system (Promega) and was measured with a Monolight 2010 luminometer. The results are expressed as the means ± SD from three independent experiments.