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. 2022 Oct 10;41(1):140–149. doi: 10.1038/s41587-022-01475-z

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — a, Relative NF-κB transcriptional reporter activity in unstimulated (left) and TNF-α-stimulated conditions (one-way analysis of variance (ANOVA) with Dunnett’s multiple comparisons test, P = 0.0395 and P = 0.0047, respectively). Error bars represent standard deviation of the mean, n = 3. b, Relative NF-κB transcriptional reporter activity at different amounts of transfected viral protein-encoding plasmid in unstimulated (top) and TNF-α-stimulated conditions (middle) (one-way ANOVA with Dunnett’s multiple comparisons test: *P = 0.0183, ****P < 0.0001 and **P = 0.0012, respectively). Error bars represent standard deviation of the mean, n = 3 (top) and n = 6 (middle). Representative anti-hemagglutinin (HA) western blot demonstrating levels of tagged viral protein in titration experiments relative to actin beta (ACTB) loading controls (bottom). a and b, Precise P values, biological repeats and n for each test are shown in Extended Data Fig. 4 and Supplementary Table 9. c, Relative NF-κB transcriptional reporter activity under unstimulated (left), TNF-α-stimulated (middle) and NSP14-induced conditions in wild-type (WT) and three independent IKBKG KO clones of HEK293 cells (two-way ANOVA with Dunnett’s multiple comparisons test). Error bars represent standard deviation of the mean, n = 3. Precise P values, biological repeats and n for each test are shown in Extended Data Fig. 4. ctrl., control. d, Schematic of viral replication assay (top) and viral replication in wild-type, mock KO and CRISPR KOs of the indicated HuSCI host targets (bottom) (Kruskal–Wallis with Dunn’s multiple comparisons test, * P = 0.031, ** P = 0.0047, *** P = 0.0003, **** P < 0.0001, respectively). Error bars represent standard deviation of the mean, n = 9. Precise P values, biological repeats and n for each test are shown in Extended Data Fig. 4 and Supplementary Table 10. e, Fluorescence microscopy images showing replication of icSARS-CoV-2-mNeonGreen in infected Vero E6 cells treated with 10 µM AZ1 or solvent (DMSO, dimethylsulfoxide). f, Cell viability and relative replication of icSARS-CoV-2-nanoluciferase in HEK293 cells (left) and Vero E6 cells (right) at different concentrations of AZ1. The EC₅₀ values were calculated with a variable slope model. Error bars represent standard deviation of the mean, n = 8 biological repeats and full analysis in Supplementary Table 11.

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