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. 2021 Oct 8;12:5907. doi: 10.1038/s41467-021-26066-5

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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 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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Fig. 7 — a Schematic of the experimental design for the perfusion of trehalose and αSyn fibril in the Brain-Chip. b Percentage of trehalose crossing the blood-brain barrier (BBB) after 24 h of perfusion. c Quantitation of the accumulated phosphorylated αSyn at the brain channel based on the fluorescence intensity at day 8 in the αSyn fibril model with or without trehalose treatment. Statistical analysis by two-way ANOVA with Tukey’s multiple comparisons test (n = 3–4 independent chips with 3–5 randomly selected different areas per chip, ***P = 0.001 compared to monomeric group, ***P = 0.0009 compared to αSyn fibrils). Error bars represent mean ± SEM. d Secreted levels of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and interferon gamma (IFN-γ) at the brain channel at D8. Statistical analysis by two-way ANOVA with Tukey’s multiple comparisons test (n = 4 independent chips with 3–5 randomly selected different areas per chip, TNF-α: **P = 0.0011 compared to monomeric group, ***P = 0.0003 compared to αSyn fibrils, IL-6: ***P = 0.0002 compared to monomeric group, ****P < 0.0001 compared to αSyn fibrils, NS not significant). Error bars represent mean ± SEM. e Quantitation of the accumulated phosphorylated αSyn at the vascular channel based on the fluorescence intensity at day 8 in the αSyn fibril model with or without trehalose treatment. Statistical analysis by two-way ANOVA with Tukey’s multiple comparisons test (n = 4 independent chips with 3–5 randomly selected different areas per chip, ****P < 0.0001). Error bars represent mean ± SEM. f Secreted levels of TNF-α, IL-6, and IFN-γ at the vascular channel at day 8 in the αSyn fibril model with or without trehalose treatment. Statistical analysis is two-way ANOVA with Tukey’s multiple comparisons test (n = 3–4 independent chips with 3–5 randomly selected different areas per chip, IL-6: **P = 0.0025 compared to monomeric group, *P = 0.0158 compared to αSyn fibrils, IFN-γ: ****P < 0.0001, NS not significant). Error bars represent mean ± SEM. g Quantitative barrier function analysis via permeability to 3 kDa fluorescent dextran at day 8 in the αSyn fibril model with or without trehalose treatment. Statistical analysis is two-sided unpaired t-test (n = 5–8 independent chips, ****P < 0.0001 compared to monomeric group, ***P = 0.0001 compared to αSyn fibrils). Error bars represent mean ± SEM. h Morphological analysis of tight junctions in endothelial cells at day 8 in the αSyn fibril model with or without trehalose treatment. The junction protein expression of Claudin-5 was visualized by immunofluorescence staining with a Claudin-5 antibody and DAPI for cell nuclei. Dashed boxes show the endothelial junctions. Scale bars: 50 μm.