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. 2022 Mar 9;10:823450. doi: 10.3389/fcell.2022.823450

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Copyright © 2022 Yaker, Tebani, Lesueur, Dias, Jung, Bekri, Guerrera, Kamel, Ausseil and Boullier.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Characterization of macrophage-derived EVs. EVs were isolated from the culture media of RAW cells incubated with (EV-LPS) or without (EV-CT) 1 µg/ml lipopolysaccharide-EK (LPS-EK) for 6 h. Evaluation of EV morphology by (A) transmission electron microscopy (TEM) and (B) cryo-electron microscopy (cryo-EM). (C) Particle-size distribution and (D) the total concentration of macrophage-derived EVs were measured by nanoparticle-tracking analysis (NTA). (E) CD9, CD81, and β-actin protein expression were assessed by western blotting. (F) Quantification of the enzymatic activity of acetylcholinesterase (AChE) in macrophage-derived EVs. Control AChE activity in the assay diluent 1X-D-PBS (CT) was defined as 100%. Data are expressed as the mean ± SEM of seven independent experiments performed in triplicate (n = 7). ***p < 0.001 vs. CT; $p < 0.05 vs. EV-CT, Mann-Whitney test.