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. 2022 Sep 13;13:1007089. doi: 10.3389/fimmu.2022.1007089

Figure 1.

Figure 1

SARS-CoV-2 Nsp14 increases NF-κB activity. (A–C) HEK293T cells were transiently transfected with V5-FLAG-Nsp14 or empty vector for 24h and then treated with or without TNF-α for 24h. V5-FLAG-Nsp14 was analyzed by protein immunoblotting (A). HEK293T cells transfected with V5-FLAG-Nsp14 or empty vector along with NF-κB-driven firefly luciferase and TK-driven renilla luciferase reporter vectors were un-treated or treated with TNF-α (B). Luciferase activity (firefly/renilla) in these cells was measured and normalized to the untreated group with the empty vector. HEK293T cells transfected with V5-FLAG-Nsp14 or empty vector were subjected to the nuclear/cytosolic fractionation. V5-FLAG-Nsp14 and NF-κB p65 in the nucleus or cytosol were analyzed by protein immunoblotting (C). Histone H3 was used as the nuclear marker. The intensity of the p65 protein band was quantified and normalized to the empty vector (D). Results were calculated from 3 independent experiments and presented as mean +/- standard error of the mean (SEM). (ns: not significant ; * p<0.05; *** p<0.001; by unpaired Student’s t-test).