Luciferase Assay for IFN-β, NF-κB, IRF3 Regulation

The luciferase assay for detecting IFN-β, NF-κB and IRF3 promoter activities were carried out as described before [21 (link)]. The plasmids pIFN-β-luc, pIRF3-luc, pNF-κB-luc, and pRL-TK were purchased from Promega. Briefly, the viral protein expression plasmids together with luciferase plasmids and pRL-TK were co-transfected into HEK293T cells. The pRL-TK vector provided constitutive expression of Renilla luciferase. After 24 h post-transfection, cells were mock- or treated with 200 ng Poly (I:C) (Sigma, China) for 12 h. The luciferase activity in cells lysates were detected using the Dual-Luciferase assay system (Promega, China) according to the manufacturer’s instructions.