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. 2021 Mar 6;19:70. doi: 10.1186/s12951-021-00801-y

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© The Author(s) 2021

Open AccessThis 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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 5 — Investigation of the gene delivery process. a Cellular internalization of LAMA-mic polyplexes in different cell lines. Cells were incubated with polyplexes of polymers and YOYO-1-labeled pDNA at N*/P 30 and analyzed via flow cytometry. Cells incubated with labeled pDNA served as control (rMFI = 1). Values represent mean ± SD (n = 3). Asterisks indicate significant differences of LPEI (a), PDMAEMA (b), LAMA-mic (c) to the control of the respective time point. b Cellular internalization of polyplexes investigated via CLSM following incubation of cells with polyplexes of polymers and YOYO-1-labeled pDNA at N*/P 30 for 1 h (n ≥ 2). Endolysosomes were stained with LysoTracker^TM Red (red), nuclei were stained with Hoechst33342 (blue) and YOYO-1 fluorescence (green) was quenched with trypan blue. Maximum intensity projection was used to display polyplexes in every plane of the cell. c Endosomal escape of LAMA-mic polyplexes was detected by the non-permeable dye calcein and analyzed via flow cytometry relative to the calcein control (rMFI = 1). Values represent mean ± SD (n = 3). If not stated otherwise, asterisks indicate significant differences to the control of the respective time point. Since there was no significant interaction of incubation time and treatment for K-562 cells, the main effects of the treatment were determined with only LAMA-mic showing a significant difference to the control (p < 0.001)