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. 2017 Jul 26;7:6494. doi: 10.1038/s41598-017-06750-7

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© The Author(s) 2017

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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Up-regulation of Notch1 expression in scratch-wounded keratinocytes. (a) After HaCaT keratinocytes were scratched, transcripts of Notch1~4 and GAPDH were determined by quantitative PCR. (b) Lysates from scratch-wounded HaCaT keratinocytes were immunoblotted with anti-NICD and anti-β-ACTIN antibodies, respectively. (c) Increased nuclear NICD expression in the region of the wound edge compared to the region far from the wound edge (outward) (>1 mm). HaCaT keratinocytes after scratching were immunostained with anti-NICD antibody. Nuclei were identified using DAPI staining. Scale bar, 25 μm. (d) Up-regulation of Hes1 promoter reporter activity after scratching. However, activity of promoter reporter containing mutated RBP-J binding sites is not up-regulated by scratching.