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. 2018 Feb 8;8:2612. doi: 10.1038/s41598-018-21000-0

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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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Expression analysis of COL4A6 and keratin markers during epithelial keratinization in vitro. hGECs were seeded in the ThinCert cell culture inserts (3D culture), and total RNA was collected after 0, 1, 3 and 7 days. mRNA expression levels of COL4A6 (A), KRT1 (B), KRT10 (C) and INV (D) were measured by real time RT-PCR. The expression of each gene was normalized to that of S29 ribosomal RNA. Bars represent the mean values and standard deviation (+/−SD) (n = 3). **p < 0.01, ***p < 0.001 (ANOVA, Tukey multiple comparison test).