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. 2018 Sep 7;8:13434. doi: 10.1038/s41598-018-31829-0

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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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Primary and SVTERT hair KC form completely differentiated skin equivalents. Normal scalp skin and HSE at day seven were analyzed by hematoxylin and eosin (H&E) and immunfluorescence staining. HSE developed a multilayered epidermis with intact stratum granulosum and stratum corneum resembling human skin. The differentiation associated proteins keratin 10, involcurin and filaggrin were detected by immunofluorescence staining, both in scalp skin and HSE cultures. keratin 2 was detected in scalp skin, but not in SE cultures. One representative experiment out of three is shown; Scale bar = 120 µm, for scalp 240 µm.