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. 2021 Mar 4;40(13):2407–2421. doi: 10.1038/s41388-021-01718-4

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© The Author(s) 2021, corrected publication 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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 7 — A Conditioned media from LNCaP parental and EnzaR cells were isolated EVs by size exclusion chromatography. Size and concentration of EVs were further analyzed by nanoparticle tracking analysis (NTA) (n = 3). B, C miR-21, COUP-TFII, and YAP1 expression levels were individually determined in EVs isolated from LNCaP parental and EnzaR cells by RT-qPCR (n = 3) and western blot. D Concentrations of EVs-isolated from sera of enzalutamide-sensitive and -resistant patients were determined by NTA analysis (n = 3). Results were presented as percentage of control. E, F miR-21, COUP-TFII, and YAP1 expression levels were individually determined in EVs isolated from sera of enzalutamide-sensitive and -resistant patients RT-qPCR (E) (n = 3) and western blot (F). G Heatmap showed the expression profiles of genes related to cancer stemness and lipid metabolism in LNCaP cells treated with EnzaR-EVs (10 μg) for 48 h. H, I LNCaP cells were treated with control (siNC), YAP1-, or CII-depleted EnzaR-EVs (10 μg) for 12 days for sphere culture (n = 3). Representative pictures and quantifications of sphere numbers were respectively shown in the H, I. J LNCaP cells treated with EnZaR-EVs derived from siNC, siYAP1, and siCII in EnzaR cells for 48 h were performed lipid staining by BODIPY 505/515 dye. K LNCaP cells were treated with control (siNC), YAP1-, or CII-depleted EnzaR-EVs (10 μg) for 48 h. Expression levels of cancer stemness-related genes such as BMI1, CDK6, TGFBR2, and SULF2 were determined by RT-qPCR (n = 3). L LNCaP cells were co-treated with enzalutamide (10 μM) and LNCaP-EVs or control (siNC), YAP1-, or CII-depleted EnzaR-EVs (10 μg) for 72 h to perform cell proliferation assays (n = 3). Asterisk indicates p < 0.05 using two-tailed Student’s t-test for above experiments.