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. 2019 Aug 15;9:11910. doi: 10.1038/s41598-019-48447-z

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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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Expression analysis of biomarker candidates for the earliest stages of ectodermal differentiation. One-step real-time RT-PCR analysis of 14 upregulated mRNAs (BTBD9, CADM1, COL9A2, DGKI, GBX2, KCNC1, KIF26B, MARCH1, PLXNA4, SLC24A2, SLC24A4, TLR4, WNT2B, and ZHX3) and one downregulated mRNA (PPP1R12B) in substate 2 of integrated analyses of mRNA and miRNA was performed using RNAs derived from substate 1 and substate 2 cells. The expression of GAPDH was used for normalization. Values are the means ± SDs (n = 3). mRNAs that showed a 1.5-fold difference in expression between substate 1 and substate 2 cells were subjected to analysis of significance using unpaired two-tailed t-tests (*P < 0.1; **P < 0.05; ***P < 0.01). Red and blue gene symbols indicate upregulated and downregulated mRNAs, respectively.