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. 2023 Feb 16;14:890. doi: 10.1038/s41467-023-36441-z

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© The Author(s) 2023

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. 5 — A, B Cholesterol depletion assays. A representative image (out of 10 randomly selected locations) for each condition is shown. Scale bars, 50 μm. A Complement was activated on CHO cells with a polyclonal anti-CHO IgG antibody. Cells were incubated with C9-depleted human serum supplemented with a chemically labeled fluorescent C9 (C9-Alexafluor 568) capable of forming MAC. B CHO cells treated with MβCD to deplete cholesterol. Complement activation and C9 detection is as described in (A). Insets show a zoomed in view of single cell. Cartoon schematics highlight the pattern of MAC deposition. C CryoEM map of the C5b9₂-CD59 complex in a lipid nanodisc applying a positive B-factor of +50 Å² (transparent surface). Surface rendering of the protein model is underlayed. CD59 is cyan, C8β is magenta and the remaining complement proteins are grey. D Map generated from the coarse-grained model of the C5b8-CD59 complex including the GPI anchor (green). Protein components colored as in panel (C). Water molecules in proximity to the membrane are shown as blue spheres. Initial and final configurations for the three MD repeats are included in the Supplementary Data Files.