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. 2021 Apr 15;11:8279. doi: 10.1038/s41598-021-87441-2

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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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Thrombospondin 1 as a novel, highly specific EV protein biomarker for neuroendocrine prostate cancer. (A) EV-associated TSP1, Gelsolin and exosomal markers CD63 and CD9 expression as analyzed by Western blotting in EVs from LNCaP-AR, LNCaP-AR-EnzR and NCI-H660 cells. (B) EV-associated TSP1, Gelsolin and exosomal markers CD63 and TSG101 expression as analyzed by Western blotting in EVs from CRPC-Adeno and CRPC-NE tissues. Clinical samples were run in two separate gels (lanes 1-9, gel no. 1 and lanes 10-18, gel no. 2). The boundaries of gels are delineated by black lines. Samples derive from the same experiment and gels/blots were processed in parallel. Following transfer of gel, blots were cut according to molecular weights. Upper parts of blots were probed with THBS1 antibody and lower parts were probed with Gelsolin and CD63 antibody. Since CD63 and TSG101 fall in same size range, following stripping, CD63 blots were re-probed with TSG101 antibody. (C) TSP1 ELISA to validate TSP1 as a novel NEPC biomarker. Left panel shows the standard curve derived from known concentrations of standard TSP1. Right panel shows computed TSP1 values from TSP1 ELISA assay using EVs from CRPC-Adeno and CRPC-NE clinical samples.