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. 2021 May 13;11:10200. doi: 10.1038/s41598-021-89655-w

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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 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/.

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Measurement of exosome secretion by qNano analysis. (A) Total number of EVs in the plasma of patients with Kidney cancer and matched controls were measured by a qNano analysis (n = 59). (B) Expression of exosome CD63, and exosome biogenesis markers Alix in the exosome of 293-T, RCC-24, 786-O, and Caki-2 cells. Exosomes were isolated by UC and filtration (0.45 μm). Full blots are presented in Supplementary Fig. S6 (C) qNano-IZON particle quantitative analysis (NP-100 nanopore) depicting a significant decrease in exosome concentrations (50–200 nm size) in the CM of RCC-24, 786-O, and Caki-2 cells treated with KTZ compared to vehicle treated controls, but not normal kidney HEK 293-T cells. *Denotes significance at p < 0.05 compared to controls and was calculated using GraphPad Prism. (D) Inhibition of exosome secretion using the MACSQuant Analyzer 10 Flow Cytometer and qNano-IZON: The exosome in the conditioned media were isolated and analyzed by the MACSQuant Analyzer 10 Flow Cytometer as described in the “Materials and methods” section. Treatments were at concentrations of 1 μM ketoconazole compounds. Exosome from DMSO treated 786-O-CD63-GFP is used as GFP control and PBS served as a reference control. *Denotes significance at p < 0.05 compared to controls and was calculated using GraphPad Prism. Data for EVs and exosomes was captured using IZON’s Control Suite software version 3.4.2.48. This software can be found at https://support.izon.com/how-can-i-get-the-latest-software-release.