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. 2021 Nov 19;12:6761. doi: 10.1038/s41467-021-26498-z

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© The Author(s) 2021

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. 2 — a Screen for SARS-CoV-2 antiviral activity of viral peptides including the G3BP inhibitor G3BPi (p = 0.000013) and N_12-27 (p = 0.000005). The amount of SARS-CoV-2 virus released was determined 16 h postinfection by focus forming assay (n = 8 independent experiments). b Affinity measurements of recombinant G3BP1 NTF2 binding to G3BPi and the SARS-CoV-2 N peptide (n = two biological duplicates each containing three technical replicates). Shown is a representative plot from one of the experiments. c Quantitative mass spectrometry comparison of G3BPi wt and ctrl purified from HeLa cells (n = 4 technical replicates). d Purification of myc-tagged SARS-CoV-2 N expressed in HeLa cells and its interaction with G3BP1 analyzed by western blot. G3BPi wt or ctrl were co-expressed with myc-tagged N where indicated. Shown is a representative blot from three independent experiments. e Effect of G3BPi on % infected VeroE6 cells during 36 h of infection (n = 9 independent experiments) *P = 0.0414. f Amount of SARS-CoV-2 RNA measured 16 h postinfection with qPCR at different MOI in VeroE6 cells expressing G3BPi or control inhibitor (n = 6 independent experiments) *P = 0.0291, **P = 0.0029, ***P = 0.0002. g Endogenous G3BP1 was purified from mock or SARS-CoV-2 infected cells and probed for N. Shown is a representative blot from three independent experiments. h In vitro interaction of recombinant full-length SARS-CoV-2 N and the NTF2 domain of G3BP1 as measured by fluorescence polarization spectroscopy (n = two biological duplicates each containing three technical replicates). Shown is a representative plot. i Quantitative mass spectrometry analysis of YFP-tagged SARS-CoV-2 N wt or 2 A purified from HeLa cells (n = 4 technical replicates). j Quantitative mass spectrometry analysis of YFP-tagged N SARS-CoV-2 and N MERS purified from HeLa cells (n = 4 technical replicates). Asterisks indicate statistical significance calculated by two-sided unpaired T test (panel a, e, f). Mean ± SD indicated throughout in the graphs. Source data are provided as a Source Data file.