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. 2020 Aug 27;9(1):1809065. doi: 10.1080/20013078.2020.1809065

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© 2020 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group on behalf of The International Society for Extracellular Vesicles.

This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 3. — Microglia cells are the main contributor to the physiological brain sEV pool. (A) Representative western blots of pooled fractions 3 and 4 of filtered EVs (sEVs; n = 4) compared to their respective total brain homogenates (TH). P2Y12 and TMEM119 were chosen as markers of microglia, PLP and CNP as markers of oligodendrocytes, synapsin1 and SNAP-25 as neuronal markers, and EAAT1 (GLAST) and EAAT2 (GLT-1) as markers of astrocytes. Of note, TMEM119 presented a lower band (at around 20 kDa) in the sEVs-enriched fractions instead of the reported 60 kDa band (approx.) observed in the TH. This could correspond to an isoform or a truncated version of TMEM119. (B) Scatter plots showing relative intensity quantifications of each cell type marker. Each lane was first referred to its total protein staining (TS) signal and the means of each group (sEVs vs. TH) were then compared in order to check for relative enrichment (with TH set to 100%). P2Y12 and TMEM119 are approximately 2.5 and 1.5 times enriched in sEVs compared to the TH, indicating a dominant contribution from microglia to the whole pool of brain sEVs. Synapsin1 and SNAP-25 are significantly decreased compared to the TH, suggesting a low contribution of neuronal sEVs to the total pool. PLP, CNP, EAAT1 and EAAT2 did not show any significant differences compared to the TH. Exact mean, SEM and p-values are given in the main text.