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. 2023 Nov 22;623(7989):1017–1025. doi: 10.1038/s41586-023-06639-8

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© The Author(s) 2023, corrected publication 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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Extended Data Fig. 1 — a, Methods used to create soluble HT1 and HT2 HIV-1 Env heterotrimers. A 20:1 transfection ratio of untagged and D7324-tagged Env expression plasmids, one of which encoded the D368R CD4 knockout mutation in gp120, was co-transfected to produce two predominant populations: untagged trimers and singly tagged trimers. Transfection supernatants were harvested and Env proteins purified by JR-52 immunoaffinity chromatography (as described), resulting in the HT1 and HT2 heterotrimers. Schematics were generated using BioRender.com. b, ELISA comparing CD4 binding of BG505, BG505 HT2, BG505 HT1, and BG505-D368R. Values from two biological replicates (n = 2) are represented by points, with the mean indicated by the dotted line.