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. 2022 Feb 22;19:53. doi: 10.1186/s12974-022-02413-1

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© The Author(s) 2022

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 5 — Endocytosis was involved in immune sensitization of monocytes induced by A53T RBC-EVs. A RBC-EVs uptake by THP-1 cells was determined by flow cytometry. Red histogram represented THP-1 cells without RBC-EVs, while blue histogram represented THP-1 cells containing RBC-EVs. The RBC-EVs were labelled with DiO. B–E Effects of endocytosis inhibitors on RBC-EVs uptake in THP-1 cells was determined by flow cytometry. Red histogram represented THP-1 cells co-treated with RBC-EVs and vehicle control, while blue histogram represented THP-1 cells co-treated with RBC-EVs and endocytosis inhibitor. F Quantitative analysis of inhibition ratio of Me-β-CD, Dynasore, Amiloride and Nocodazole. G Levels of oligomeric α-syn in THP-1 cells pretreated with A53T RBC-EVs, along with Me-β-CD, Nocodazole or Amiloride. H Representative confocal image of THP-1 cells incubated with DiI (red)-labelled RBC-EVs after treatment with Dynasore, Nocodazole Me-β-CD and Amiloride, cell nucleus was labelled by Hoechst 33258. Scale bar, 10 µm. I Quantitative analysis of pro-inflammatory cytokines IL-1β, IL-6, TNF-α, IL-8, IFN-γ, IL-2, and IL-12p70, and J anti-inflammatory cytokines IL-4, IL-10, and IL-13 using MSD, released by THP-1 cells pretreated with A53T RBC-EVs alone or with Nocodazole or Me-β-CD, followed by LPS stimulation. N = 3 independent experiments in each group. Values are means ± S.E.M, one-way ANOVA test. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001