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. 2019 Mar 13;10:1203. doi: 10.1038/s41467-019-09174-1

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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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Fig. 4 — Runx2 acts as a mediator of the pro-calcifying effects of PARP1. a Western blot analysis of several osteogenic factors expression in calcified VSMCs transfected with Scr or PARP1 shRNA. (n = 5 per group). b The expression levels of Runx2 and SMA in abdominal arteries of indicated groups were determined by immunofluorescence staining. (n = 5 per group). Scale bar, 100 μm. c–e Rat abdominal aortas were inoculated with Ad-Scr shRNA, or Ad-Runx2 shRNA together with Ad-Null or Ad-PARP1 at three weeks after the adenine diet. Six weeks later, arteries were isolated and the calcification was analyzed by H&E, von Kossa staining (c) and the calcium quantification (d) (n = 10–12 per group), and the downstream osteogenic markers (OPN, ColIA1, OC, and Runx2) were analyzed by western blot (e). (n = 5 per group). Scale bar, 100 μm. f, g The aortic rings (f) or rVSMCs (g) were pre-infected with Ad-Scr shRNA, or Ad-Runx2 shRNA together with Ad-Null or Ad-PARP1, and exposed to the osteogenic medium for 14 days. The calcification was determined by the calcium assay respectively. (n = 5 for each group). Statistical significance was assessed using one-way ANOVA for multiple comparison and is presented as follows: NS: no significance, **P < 0.01 and ^##P < 0.01. All values are means ± S.D. Source data are provided as a Source Data file