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. 2025 Jan 2;16:250. doi: 10.1038/s41467-024-55531-0

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© The Author(s) 2025

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, 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 you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. 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-nc-nd/4.0/.

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Fig. 1 — a Procedure of the single-molecule SNARE disassembly assay on total internal reflection fluorescence (TIRF) microscope. b TIRFM images of Cy3 fluorescence channel containing SNARE-nanodisc after immobilisation along with a pictorial representation of nanodisc present on the glass slide before and after the disassembly by NSF. c Number of Stx-Cy3 nanodiscs present under various experimental conditions. Disassembly reaction in the presence of all components (brown), without the addition of Mg²⁺ (light grey), only with ATP already bound to the NSF (No extra ATP) and other controls. p-values calculated using a two-tailed paired t test and mean ± SD values are given, ***= p < 0.005. (n = 3-6 independent SNARE disassembly experiments).