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. 2025 Jan 24;16:996. doi: 10.1038/s41467-024-55137-6

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© The Author(s) 2025, corrected publication 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. 5 — a, b Artificial endosomes were constructed via thin film hydration of DOPE, DOPC, DOPS, NBD-PE, and Liss-Rhod-PE at a molar ratio of 48:25:25:1:1, respectively. Endosomes were mixed with a C8-200, E4i-200, E4t-200, and E4s-200 (C8/E4x 200-core) LNPs or b C12-200, E8i-200, E8t-200, and E8s-200 (C12/E8x 200-core) LNPs, and fluorescence was measured at 465/520 nm at several timepoints over 24 h. Relative fluorescence is reported as mean ± SD of n = 3 technical replicates. Atomistic MD simulations were performed by modeling the insertion of a single IL into a lipid bilayer membrane with a mixture of DOPE, DOPC, and DOPS at a molar ratio of 2:1:1, respectively for c C8/E4x 200-core ILs and d C12/E8x 200-core ILs. The vertical distance of the center of mass of the IL to the bilayer center is reported as mean ± SD of n = 3 technical replicates. Correlation between artificial endosomal rupturing and bilayer penetration experiments for e C8/E4x 200-core LNPs/ILs and f C12/E8x 200-core LNPs/ILs. Data is represented as mean ± SD of bilayer penetration (horizontal) and endosomal disruption (vertical) of n = 3 technical replicates. Snapshots during the simulations of g C8/E4x 200-core ILs and h C12/E8x 200-core ILs. The IL is colored magenta and the nitrogen, oxygen, carbon, and hydrogen atoms in the lipid bilayer are colored blue, red, white, and white, respectively. Two-way ANOVA with Holm–Šídák correction for multiple comparisons were used to compare relative fluorescence across timepoints and LNPs to a C8-200 and b C12-200. The least significant adjusted p value out of the three branching groups is shown above each timepoint. One-way ANOVA with Holm–Šídák correction for multiple comparisons were used to compare distance to bilayer center across treatment groups to c C8-200 and d C12-200. Source data are provided as a Source Data file.