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. 2021 Jun 29;11:13471. doi: 10.1038/s41598-021-92860-2

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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 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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Cucurbitacin B, gossypol, and obatoclax induced EV secretion in a variety of cells. (a–c) HEK293T cells were treated with 1 μM amlodipine, 0.3 μM osimertinib, 1 μM cucurbitacin B, 1 μM doramectin, 0.3 μM gossypol, 10 μM HA14-1, 10 μM miltefosine, or 0.03 μM obatoclax for 24 h and then secreted EVs were determined using a TIM4-CD63 ELISA (a), TIM4-CD9, TIM4-CD63, TIM4-CD81 ELISA (b) or NTA (c). (d) Cell lines were treated with cucurbitacin B, gossypol, or obatoclax at the concentrations shown on Table 3 for 24 h, and then secreted EVs were determined using a TIM4-ELISA. EVs secreted from hMSC, K562, HCT116, SW480, SW620, and NIH/3T3 cells were detected using TIM4-CD63 ELISA. EVs secreted from THP-1, EL4, and Jurkat cells were detected using TIM4-CD81 ELISA. *p < 0.05, **p < 0.01, versus DMSO, Student’s t-test.