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. 2019 Jul 24;9:10739. doi: 10.1038/s41598-019-47189-2

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© The Author(s) 2019

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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Expression of AR-FL and AR-V7 in an isogenic model of enzalutamide resistance. A standard curve method using a qPCR-based assay was used to determine the copy number of (A) AR-FL and (B) AR-V7. While AR-FL was consistent across all cell lines, AR-V7 varied according to enzalutamide resistance status. Data graphed as mean ± SEM (n = 3). Statistical analysis performed using ordinary one-way ANOVA (***p ≤ 0.001). RNA in situ hybridisation was used to determine the expression of (C) AR-FL and (D) AR-V7 in FFPE cell plugs. Positive expression was confirmed by the presence of punctate red staining (as indicated by arrow). No differences were observed for AR-FL, however AR-V7 expression was prominent in LNCaP clone 1 compared to the other two cell lines. PC-3 cells were used as a negative control. Representative images are shown at 40x magnification.