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. 2021 Oct 6;11:19896. doi: 10.1038/s41598-021-99180-5

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© The Author(s) 2021

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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CLSM of cells treated with protein-encapsulating LNPs and the percentages of cells having cytosolic/nuclear localization of the proteins 6 h after treatment with the following: (A) NLS-EGFP alone (left) and NLS-EGFP-LNPs (1:10) (right); (B) NLS-(− 30)GFP alone (left), NLS-(− 30)GFP-LNPs (1:10) (PEG(–)) (center), and NLS-(–30)GFP-LNPs (right); (C) NLS-(− 30)GFP-LNPs (1:10) (DOPE) (center), and NLS-(–30)GFP-LNPs). The percentages of cells bearing cytosolic NLS-EGFP/NLS-(− 30)GFP signals are presented as the mean ± SD (n = 3). n.s., not significant; ***P < 0.001 (one-way analysis of variance (ANOVA) followed by Tukey–Kramer’s honestly-significant difference test). Protein concentration, 2.5 µM. Concentration of LNPs were set to yield 2.5 µM NLS-EGFP or NLS-(− 30)GFP when hypothesized that all the employed proteins for LNP preparation was encapsulated into the LNPs.