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. 2018 Jan 31;8:1949. doi: 10.1038/s41598-018-19458-z

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© The Author(s) 2018

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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Relative quantification of PCA3 expression depends on mRNA reference gene. Cells were cultured using RPMI 1640 medium containing 10% charcoal-stripped FBS for 24 hours prior to treatment with different DHT concentrations. The cDNA was prepared from these cells for the amplification of PCA3 and mRNA reference genes by qPCR. Relative expression of PCA3 across all cell lines was normalised by the best reference genes combination (ACTB-GAPDH) (A), by the most stable single gene ACTB (B) or GAPDH (C), by the least stable single gene 18 S rRNA (D), or K-ALPHA-1 (E), or ALAS1 (F). Y-axis indicates the fold change of relative expression of PCA3 in RWPE-1 cells set to 1. Error bars indicate the standard error (±SE) evaluated from three biological replicates. * and **Indicate P < 0.05 and P < 0.01, respectively.