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. 2019 Dec 6;9:18534. doi: 10.1038/s41598-019-54582-4

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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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Ex vivo expression analysis of Bd reference gene candidates. Shown is the variation in the mRNA expression (Cq) of GAPDH, TUB, R6046 and APRT in (a–c) 2 × 10⁷ Bd spores and (d–f) 5 × 10⁷ Bd spores that were incubated for 2 hours at 20 °C (c,f) with or (b,e) without skin tissue of midwife toads, or (a,d) a combination of both. Every condition was tested in fourfold. The whiskers represent the median, the minimum and maximum values, and the first and third quartiles. A plus (+) indicates the mean Cq value. (g) GeNorm stability analysis providing a GeNorm M and coefficient of variation (CV) value, combined with NormFinder intergroup variation analysis.