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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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Extended Data Fig. 5 — a, Y2H_HIS3 yeast growth on selective plates of HuSCI interaction partners as DB-fusion proteins tested against AD-fusion of the SARS-CoV-2 Nucleocapsid (AD-N) protein (Wuhan-Hu1, original screen) and AD-N containing ‘lineage defining’ amino acid substitutions: D3L and S235F (α-strain), T205I (β-strain), or P80R (γ-strain). Shown is one representative result of 5 repeats. b, Y2H_HIS3 yeast growth on selective plates of HuSCI interaction partners as DB-fusion proteins tested against AD-fusion of the SARS-CoV-2 Envelope (AD-E) protein (Wuhan-Hu1, original screen) or AD-E containing ‘lineage defining’ substitution P71L (β-strain). Shown is one representative Y2H_HIS3 result on selective media, out of 2 repeats. a - c, Black circles indicate changes in yeast colony growth between human proteins tested against viral variant ORFs or the originally screened Wuhan strain ORFs observed consistently across all repeats. c, AD-empty control plate for a, b indicates lack of autoactivation. d, Layout of DB-fusion HuSCI interactors (purple) tested with AD fusion SARS-CoV-2 proteins or AD-empty control, respectively in a - c. N/A indicates human interactors.