Figure 1.
SARS-CoV-2 N protein inhibits NF-κB pathway, unlike SARS-CoV N protein. (A and B) HEK293 cells were transfected with NF-κB-Luc, along with plasmids encoding the indicated Flag-SARS-CoV-2 proteins. At 24 h after transfection, cells were infected with SeV (A) for 10 h or stimulated by IL-1β (B) for 4 h, and luciferase activity was measured. (C and D) HEK293 cells were transfected with NF-κB-luc (C) or IL-8-luc (D), along with increasing amounts of plasmids encoding Flag-SARS-CoV-2 N protein. Cells were infected with SeV for 10 h and assayed for luciferase activity. (E and F) HeLa cells were transfected with the plasmid encoding Flag-SARS-CoV-2 N protein for 24 h, cells were infected with SeV (E) for 8 h or stimulated by IL-1β (F) for 4 h, and total RNA extracted from the cells was evaluated by RT-qPCR. (G) A549 cells were transfected with the plasmid encoding Flag-SARS-CoV-2 N protein at 24 h after transfection; cells were stimulated by TNF-α for 0, 1, 2 h. The lysates were harvested for immunoblot analysis. (H and I) HEK293 cells were transfected with NF-κB-Luc (H) or IL-8-Luc (I), along with increasing amounts of plasmids encoding Flag-SARS-CoV N protein. Cells were infected with SeV for 10 h and assayed for luciferase activity. (J and K) HeLa cells were transfected with the plasmid encoding Flag-SARS-CoV N protein for 24 h, cells were infected with SeV (J) for 8 h or stimulated by IL-1β (K) for 4 h, and total RNA extracted from the cells was evaluated by RT-qPCR. (L) A549 cells were transfected with the plasmid encoding Myc-SARS-CoV N protein, at 24 h after transfection, cells were stimulated by TNF-α for 0, 1, 2 h. The lysates were harvested for immunoblot analysis. Horizontal lines in figures represent the average value of the positive control group. Graphs show mean ± SEM (n = 3 in A–F and H–K) from one representative experiment. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, ns, not significant (unpaired, two-tailed Student’s t test). Data in G and L are done at least twice, and one representative is shown. Data in A–F and H–K are done in at least three independent experiments. Source data are available for this figure: SourceData F1.

SARS-CoV-2 N protein inhibits NF-κB pathway, unlike SARS-CoV N protein. (A and B) HEK293 cells were transfected with NF-κB-Luc, along with plasmids encoding the indicated Flag-SARS-CoV-2 proteins. At 24 h after transfection, cells were infected with SeV (A) for 10 h or stimulated by IL-1β (B) for 4 h, and luciferase activity was measured. (C and D) HEK293 cells were transfected with NF-κB-luc (C) or IL-8-luc (D), along with increasing amounts of plasmids encoding Flag-SARS-CoV-2 N protein. Cells were infected with SeV for 10 h and assayed for luciferase activity. (E and F) HeLa cells were transfected with the plasmid encoding Flag-SARS-CoV-2 N protein for 24 h, cells were infected with SeV (E) for 8 h or stimulated by IL-1β (F) for 4 h, and total RNA extracted from the cells was evaluated by RT-qPCR. (G) A549 cells were transfected with the plasmid encoding Flag-SARS-CoV-2 N protein at 24 h after transfection; cells were stimulated by TNF-α for 0, 1, 2 h. The lysates were harvested for immunoblot analysis. (H and I) HEK293 cells were transfected with NF-κB-Luc (H) or IL-8-Luc (I), along with increasing amounts of plasmids encoding Flag-SARS-CoV N protein. Cells were infected with SeV for 10 h and assayed for luciferase activity. (J and K) HeLa cells were transfected with the plasmid encoding Flag-SARS-CoV N protein for 24 h, cells were infected with SeV (J) for 8 h or stimulated by IL-1β (K) for 4 h, and total RNA extracted from the cells was evaluated by RT-qPCR. (L) A549 cells were transfected with the plasmid encoding Myc-SARS-CoV N protein, at 24 h after transfection, cells were stimulated by TNF-α for 0, 1, 2 h. The lysates were harvested for immunoblot analysis. Horizontal lines in figures represent the average value of the positive control group. Graphs show mean ± SEM (n = 3 in A–F and H–K) from one representative experiment. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, ns, not significant (unpaired, two-tailed Student’s t test). Data in G and L are done at least twice, and one representative is shown. Data in A–F and H–K are done in at least three independent experiments. Source data are available for this figure: SourceData F1.

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