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. 2018 Jan 10;8:253. doi: 10.1038/s41598-017-18050-1

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© The Author(s) 2017

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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Monolayer and micro-tumour behaviour of C42B and LNCaP cell lines in androgen deprived conditions. (a) C42B (Top) and LNCaP (Bottom) cells were seeded in expansion culture medium for 24 hours followed by medium exchange to androgen-depleted medium (CSS) for a further 48 hours. Abiraterone Acetate was then added to the culture medium at the indicated concentrations for an additional 48 hours. AlamarBlue, Cell Titre-Glo 3D Cell Viability and PicoGreen assays were then performed to assess metabolic activity, ATP quantity, and DNA quantity, respectively. All results are represented as a percentage of the FBS-containing culture medium control values (data was collected from three independent experiments, each having four technical replicate cultures). Statistical significance was calculated by two-way ANOVA compared to the corresponding zero value (**P < 0.0001 and *P < 0.01) or compare 2D and 3D values with the same drug treatment (Ψ P < 0.005). (b) Metabolic activity (AlamarBlue assay) and DHR123 staining of LNCaP monolayers at specified Abiraterone Acetate concentrations. Results represented as the mean fluorescence values of four individual samples normalized to control culture values. Statistical significance was performed using one-way ANOVA (*** P<0.001). Side panel represents the cellular morphology of LNCaP cells at the indicated Abiraterone Acetate concentrations (µM). Scale bar = 200 µm.