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. 2024 Sep 5;14:20697. doi: 10.1038/s41598-024-71305-6

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© The Author(s) 2024

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 5 — (a) Activation of the assay by WT Nsp5 and the C145A catalytically inactive mutant 20 h post transfection. Mean of 3 independent experiments + SD. (b) Representative Western blot showing the expression levels of Nsp5 catalytic mutant and its inability to cleave ACE2 reporter. (c) Gaussia activity induced by WT Nsp5 protein and E166V Nsp mutant in the presence of nirmatrelvir and (d) relative sensitivity of both proteins to the inhibitor. Mean of 3 independent experiments + SD. ***P < 0.001, unpaired Student’s t-test.