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. 2021 Feb 9;11:3438. doi: 10.1038/s41598-021-82901-1

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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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Astrocytic hApoEε4 alters the dynamics of cholesterol and tau oligomer accumulation. Scale bar of representative images represents 50 µM. High-content imaging and its associated quantification represents data from one independent experiment. (a) Representative live-cell imaging of GFP-labeled cholesterol internalization one day following a 4-h incubation in co-cultures. Hoechst stain was used to visualize nuclei. (b) Quantification of Hoechst-labeled nuclei (n = 6 wells/group) in panel (a). (c) Quantification of the total GFP (cholesterol) intensity in the cytoplasm of cells by high-content image analysis. Data are depicted as fold change over P_GFAP::hApoEε3 group (n = 6 wells/group). **p < 0.01, ***p < 0.005, Tukey’s multiple comparisons test following one-way ANOVA. (d) Representative live-cell imaging of cy3-tagged tau internalization 24 h following cy3-tau addition. Hoechst stain was used to visualize nuclei. (e) Quantification of Hoechst-labeled nuclei by high-content imaging (n = 5 wells/group). *p < 0.05, **p < 0.01, Tukey’s multiple comparisons test following one-way ANOVA. (f) Quantification of the number of cy3-positive cells. Data are depicted as the fold changes over respective monomer or oligomer-treated P_GFAP::EV groups (n = 5 wells/group). NS not significant, *p < 0.05, Tukey’s multiple comparisons test following one-way ANOVA. (g) Representative live-cell imaging of cy3-tagged tau internalization 24 h following tau addition. Cells were pre-treated with vehicle or 150 mU heparinase III for four hours prior to cy3-tau oligomer addition. Hoechst stain was used to visualize nuclei. (h) Quantification of Hoechst-labeled nuclei by high-content imaging (n = 3 wells/group). (i) Quantification of the number of cy3-positive cells. Data are depicted as the fold change over P_GFAP::hApoEε3 + vehicle group (n = 3 wells/group). NS not significant, **p < 0.01, Fisher’s LSD test following one-way ANOVA.