FIG 7.

NS6 is not an RNA binding protein. HEK-293T cells were transfected with plasmids encoding Flag-tagged RIG-I protein, PDCoV NS6 and N protein, or empty vector, respectively. Cells were lysed 28 h after transfection, and the resulting supernatants were incubated with poly(C)- or poly(I·C)-coated agarose beads for 4 h at 4°C. The beads were then washed three times with lysis buffer by centrifugation, followed by Western blotting with mouse anti-Flag antibody. The poly(I·C)-coated agarose beads (pIC-beads) were prepared from poly(C)-coated beads (pC-beads; Sigma) by incubating them with an equal volume of 2 mg of poly(I) (Sigma) per ml for 1 h at 56°C.