Figure 3.

SADS-CoV N protein suppresses the RLRs signaling pathway. (A–F) pCMV-FLAG-N and a series of expression plasmids encoding the RLRs’ signaling pathway adaptors (RIG-I, MDA5, IPS-1, TRAF3, IKKϵ, TBK1, and IRF3), together with a luciferase reporter plasmid IFN-β-Luc and the internal control plasmid pRL-TK were co-transfected to HEK-293T cells. After 24 h, cell lysates were collected using a passive lysis buffer, and the relative activity of the IFN-β promoter was measured with the Dual-luciferase assay. Or cell protein lysates were collected using RIPA lysis buffer, and the expression of SADS-CoV N protein and β-actin (as the loading control) was detected by Western blot. **P < 0.01; ns, nonsignificant differences in data.