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. 2021 Feb 19;13:593927. doi: 10.3389/fnagi.2021.593927

FIGURE 5.

FIGURE 5

Postmortem Human CNT, SAD and FAD astrocytes in culture and characterization of AD A-EVs. (A) Representative images show astrocyte cultures after 50 days. Magnification × 4, scale bar 40 μm. White arrow indicates focal cell aggregates in FAD culture. (B) Morphological characterization showing the following: F-actin visualized in red, GFAP visualized in green and nuclei in blue. Magnification × 20, scale bar 100 μm, magnification × 60, scale bar 10 μm. (C) MFI of GFAP data seen in B. 20X Representative data from CNT, n = 2; SAD, n = 3; FAD, n = 3. (D) Concentration of EVs/mL measured by flow cytometry. (E) Concentration of EVs/mL measured In the different size intervals: MVs, 0,1-1 μm; Apoptotic bodies, 1-6 μm. (F) Heatmap of percentages of EVs positive for cell markers (AQ4, astrocyte; Annexin, phosphatidylserine (PS) and DIOC6, mitochondria) from each CNT, FAD, and SAD. (G) Comparative of percentages of EVs positive of cell markers for grouped data seen in panel (F). (H) Heatmap of the normalized MFI of cell markers (to mean data from CNT individuals). (I) Comparative of MFI of cell markers for grouped data seen in panel (H). Representative data from CNT, n = 3; SAD, n = 3; FAD, n = 3. Data was plotted as means and SEM. One-way ANOVA, Tukey’s multiple comparison test. ∗Indicates p < 0.05; ∗∗ indicates p < 0.01.