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. 2022 Aug 25;14:933979. doi: 10.3389/fnagi.2022.933979

FIGURE 5.

FIGURE 5

Aβ42 oligomers (Aβ42) oligomers and misfolded tau increase small EV secretion and are associated with secreted small EVs. (A) Representative images of DiI-stained small EVs secreted from HT22 cells with or without 250 nM Aβ treatment for 24 h. (B) Quantification of secreted small EVs for (A). Data are presented as means ± SEM. n = 3 independent experiments, t-test, **P < 0.005. (C) Representative immunoblots for the indicated proteins in secreted small EVs and cell lysates from HT22 cells with or without 250 nM Aβ treatment for 24 h. (D) Quantification of Hsc70 and TSG101 (small EV markers) for (C). Data are presented as means ± SEM. n = 4 independent experiments, t-test, ***P = 0.0007, **P < 0.004. (E) Representative NTA analysis of vesicle size distribution of small EVs isolated from HT22 cells. The black line indicates concentration and size of particles, and the red line indicates standard error. (F) Representative images of DiI-stained small EVs secreted by DIV10 cortical primary neurons derived from C57BL6 (WT) or tauP301S (P301S) mice. (G) Quantification of secreted small EVs for (F). Data are presented as means ± SEM. n = 3 independent experiments, t-test, **P = 0.002. (H) Representative immunoblots for the indicated proteins in secreted small EVs and cell lysates from tet-inducible iHEK-P301L cells expressing tauP301L, with or without tetracycline. (I) Quantification of Hsc70 and TSG101 (small EV markers) for (H). Data are presented as means ± SEM. n = 4 independent experiments, t-test, ***P < 0.0003.