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. 2019 Sep 11;9:13129. doi: 10.1038/s41598-019-49474-6

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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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Optimal number of reference genes for normalization of RT-qPCR data suggested by geNorm. The tool calculates the pair-wise variation V_n/V_n + 1, where n is the number of reference genes used for normalization. According to Vandesompele and co-workers⁴⁶, V value below 0.15 (dotted line) signifies that inclusion of an additional gene (n + 1) to the normalization factor yields no benefit.