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. 2018 May 4;9(1):818–836. doi: 10.1080/21505594.2018.1451184

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© 2018 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group

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

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Figure 8. — Evaluation of cell death upon treatment of CHO and T98G cells with EVs or EVs-free supernatant. Cells were stained with the Annexin V-FITC (green)/ PI (orange) kit, fluorescence images were recorded and the intensity of cell analyzed for each channel. (A-D) CHO evaluation upon treatment with A. castellanii EVs. (A) untreated CHO control, (B) saponin treated CHO, (C) EVs treated and (D) EVs-free supernatant treated CHO cells. (E-H) T98G treatment with A. castellanii EVs. (E) untreated T98G cells, (F) saponin treated T98G, (G) EVs treated and (H) EVs-free supernatant treated CHO cells. Pictures bellow each graph are representative of the microscopy images. Saponin treated CHO or T98G cells, and CHO treated with EVs or EVs-free supernatant displayed double positivity (PI⁺/Annexin V⁺), with strong stain of nuclei (orange) and phosphatidylserine on cell membrane (green), suggesting a necrotic process. T98G cells treated with EVs or EVs-free supernatant displayed a single stain (PI⁻/Annexin V⁺), suggesting apoptosis.