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. 2019 Jun 3;9:8193. doi: 10.1038/s41598-019-44668-4

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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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Medial calcification is induced in carotid artery after vitamin D₃ injection. Male SD rats were injected with vehicle or 5 mg/kg vitamin D₃ (VitD₃) for 3 consecutive days, and carotid arteries were harvested 14 days later. (A) Von Kossa staining for calcium deposits. Arrows show calcium deposits in medial layer. (B) Calcium content was measured in the carotid arteries from vehicle (n = 7) or vitamin D₃-injected rats (n = 11) using the o-cresolphthalein complexone method. (C) Vitamin D₃ levels in serum were measured at 14 days in vehicle (n = 5) and vitamin D₃-injected (n = 6) rats. Values are mean ± SEM. *P < 0.05.