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. 2020 Dec 30;13(1):47. doi: 10.3390/v13010047

Figure 2.

Figure 2

SARS-CoV-2 N protein represses IFN-β production by targeting the RIG-I pathway. (A–E) A549 cells were co-transfected with pHA-N, together with pFlag-RIG-I (A) or pFlag-MAVS (B), pFlag-TBK1 (C), pFlag-IKKε (D), of pFlag-IRF3/5D (E) for 24 h. Cell lysates were harvested, IFN-β mRNA was determined by q-PCR (top), and HA-N, Flag-RIG-I (A), MAVS (B), TBK1 (C), IKKε (D), and pFlag-IRF3/5D (E) were confirmed by Western blotting (bottom). (F–J) HEK293T cells were co-transfected with pIFN-β-Luc, pPRL-TK, and pHA-N, together with pFlag-RIG-I (F), pFlag-MAVS (G), pFlag-TBK1 (H), pFlag-IKKε (I), and pFlag-IRF3/5D (J) for 24 h. Cell lysates were harvested, IFN-β-Luc reporter activity was determined by dual luciferase reporter assays (top), and HA-N, Flag-RIG-I (F), or MAVS (G), TBK1(H), IKKε(I), and pFlag-IRF3/5D (J) were confirmed by Western blotting (bottom). Data in A–F were expressed as the mean ± SEM of at least three independent experiments. ns, there was no significant difference, ** p < 0.01, *** p < 0.001.