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. 2020 Feb 20;10:3087. doi: 10.1038/s41598-020-59926-z

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© The Author(s) 2020

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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Time-lapse micrographs of the lipid-mixing assay between lipopeptide-functionalized GUVs and lipopeptide-functionalized LUVs before (time = 0 minutes) and after (time = 10, 30 and 60 minutes) appearance of LUVs in the confocal volume. The GUVs are excited at 488 nm and fluorescence emission is detected between 500–550 nm (green), while LUVs are excited at 633 nm and emission is detected between 650–700 nm (blue). The lower panels show a selected GUV magnified at time = 0, 5, 10, 15, 30 and 60 minutes during the time-lapse lipid-mixing experiment and the corresponding fluorescence intensity evolution along the cross-section of the lipid bilayer is plotted in the graphs. (A) Lipid-mixing assay between CP₄K₄-functionalized GUVs and CP₄E₄-functionalized LUVs, (B) lipid-mixing assay between CP₄E₄-functionalized GUVs and CP₄K₄-functionalized LUVs, (C) lipid-mixing assay between non-functionalized GUVs and non-functionalized LUVs. Imaging was performed every minute for one hour using a Leica TCS SPE microscope.