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. 2018 Jul 9;8:10332. doi: 10.1038/s41598-018-28742-x

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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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Schematic representation of the methods of pixel analyses (a,b) and the practical images used in the analyses (c). (a) Hallmark cells were set in the corner of the region of interest (ROI) at time point 0 hours; an area of ROI at time point 0 hours was set about 6,000 pixels. The ROI at each time point was cropped based on the hallmark cells. (b) Binarization processing using MetaMorph. Cell areas were set as white pixels and extracellular matrixes as black pixels. (c) Practical images used in pixel analyses in the proliferative (upper panels) and prehypertrophic-hypertrophic (lower panels) zones. In series of pictures in each zone, vehicle-treated (upper pictures) and CNP-treated (lower pictures) are indicated at each time point. Yellow dashed quadrangle indicate the ROIs.