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. 2018 Aug 10;8:11990. doi: 10.1038/s41598-018-30463-0

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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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Real-time quantitative polymerase chain reaction-based validation of key genes associated with efferent and afferent nerve fibre development. Heat map clustering displaying the relative mRNA expression levels of 9 differentially expressed genes selected from all 24 samples (8 groups in triplicate). The orange colour indicates the genes that are upregulated, and the blue colour indicates the genes that are downregulated. The values were calculated using comparative 2^−ΔΔCt method, with Gapdh levels as the internal control (efferent nerve fibres are shown in yellow and afferent nerve fibres are shown in purple). Error bars represent standard deviations from the mean. Data were obtained in three independent experiments. ^{*, #,§}p < 0.05; ^{**, ##, §§}p < 0.01; ^*in efferent nerve fibre development, ^#in afferent nerve fibre development, ^§difference between efferent and afferent nerve fibre expression.