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. 2023 Jul 4;14:3945. doi: 10.1038/s41467-023-39693-x

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© The Author(s) 2023

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. 3 — a Diagram of migrasome collection and treatment to endothelial cells. b Left: representative TEM images of the isolated migrasomes after negative staining. Middle: diameter of PBS-M and Aβ40-M. Right: Zeta potential of the isolated migrasomes. Data are representative of 3 biologically independent experiments. Lines in plots represent mean values. c Expression of migrasome markers in the purified migrasomes was assessed with western blot. Data are representative of 3 biologically independent experiments. d–g Migrasomes were treated to endothelial cells at a concentration of 50 μg/ml for 2 h. d Frequency of endothelial cells (green) attached with migrasomes (red) and the number of migrasomes bond to each endothelial cell were calculated. Data are representative of 5 biologically independent experiments. Diameter of cluster for PBS-M (N = 201) and Aβ40-M (N = 160) was also quantified. Data are presented as mean values ± SEM with the indicated significance (by two-tailed Student’s t test). e Endothelial cells were treated with PBS-M, Aβ40-M or Aβ42-M then subjected to SYTO10 and Ethidium homodimer-2 staining. The number of intact cells (SYTO10⁺Ethidium homodimer-2^-) and injured/dead cells (SYTO10⁺Ethidium homodimer-2⁺) was calculated. Data are representative of 3 biologically independent experiments. Left: representative images showing the cytotoxic property. Middle: Comparison of injured/dead cells in endothelial cells treated with PBS-M, Aβ40-M or Aβ42-M. Right: Percentage of intact cells (grey bars) and injured/dead cells (red bars) in PBS-M-, Aβ40-M- or Aβ42-M-treated endothelial cells. Data are presented as mean values ± SEM with the indicated significance (compared with MOCK by one-way ANOVA followed by Tukey’s post-test). f–g Endothelial cells were treated with 50 μg/ml of PBS-M or Aβ40-M for 2 h then subjected to bulk RNAseq. N = 2 in each group. f Gene Ontology-biological process (GO-BP) enrichment of the downregulated genes in Aβ40-M treated endothelial cells. g Heatmap showing the transcription of tight junction protein related genes of PBS-M and Aβ40-M treated endothelial cells. Source data are provided as a Source Data file.