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. 2022 May 4;13:893692. doi: 10.3389/fmicb.2022.893692

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Copyright © 2022 Rada, Hrdý, Zdrha, Narayanasamy, Smutná, Horáčková, Harant, Beneš, Ong, Tsai, Luo, Chiu, Tang and Tachezy.

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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Small EVs produced by Trichomonas vaginalis TV79-49c1⁺ contain Trichomonasvirus (TVV) particles. (A) Immunoblot analysis of 24 fractions collected from the last step of EV separation using linear OptiPrep gradient. Antibodies against Tsp1 (sEV marker), and TVV1 and TVV3 capsid proteins were used for screening of fractions. (B) Densitometry of western blots. (C) Protein protection assay. Membrane protection of TVV proteins was monitored by immunoblotting using antibodies against TVV1 capsid protein. P1, the pool of fractions 10–12; P2, the pool of fractions 17–19; T, trypsin; Tx, Triton X100. (D) Multi-angle dynamic light scattering analysis of sEV size in fractions 17, 18, and 19. (E) (a–c) Cryo-electron microscopy of sEV fraction. (F) (a–f). Electron microscopy of sEVs after negative staining. Arrows point to TVV particles. Bars indicate 40 nm.