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. 2021 Jan 6;12(1):23. doi: 10.1038/s41419-020-03310-2

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© The Author(s) 2021

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. 2 — a The procedure of primary neuronal-conditional media (NCM) collection and coculture with primary microglia. b Positive rate of Rat primary cortical neuron (MAP2) and microglia (IBA1). c The relative miR-98 expression level of microglia after added with NCM from starvation and OGD-R treatment and GW4869 blocking. (n = 3 independent experiments). d The relative miR-98 expression level between neuron and microglia of purification culture, using the miR-98 expression of Neuro-2a (N2a) cells as a baseline. e The procedure of extraction of primary neuronal EVs and co-cultured with microglia. f Transmission electron micrograph (TEM) of neuronal EVs. g Size distribution of neuronal EVs based on Zetasizer Nano-ZS90 measurements. h Western blot for EVs markers Alix, TSG101, and Calnexin. i Representative images of internalization of PKH26-labeled neuronal EVs into the CellTracker^TM Green-labeled microglia 12 h after coculture. j Relative miR-98 level in microglia 24 h after incubated with PBS and neuronal EVs (extraction by ~80~100 ml neuronal media). (n = 5 independent experiments). Data are shown as means±S.E.M., ^*P, ^#P < 0.05; ^**P < 0.01; ^***P < 0.001 vs corresponding control. k–n Transmission electron micrograph (TEM) of serum EVs from both individuals and rats. o–p Size distribution of serum EVs from both individuals and rats. q Western blot for EVs markers Alix, TSG101, and GAPDH. r The relative miR-98 expression level between in serum EVs between both individuals and rats.