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. 2018 Jan 12;8:658. doi: 10.1038/s41598-017-19092-1

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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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(A) Time course of IL-24 mRNA expression levels induced by iron, TNF-alpha or both iron and TNF-alpha stimulation. The time course of IL-24 gene expression was evaluated by real-time PCR on days 1, 3, 6, 9, and 12 after the addition of 100 µg/mL iron (holo-transferrin) and/or 1 ng/mL TNF-alpha to the calcification medium. The gene IL-24 expression level was enhanced by iron and TNF-alpha stimulation, and the increased gene expression level was maintained during the cell culture period. These experiments used one cell lines of HASMCs. (B) Time course of IL-24 by ELISA induced by iron, TNF-alpha or both iron and TNF-alpha stimulation. The time course of IL-24 protein expression was evaluated by real-time PCR on days 1, 3, 6, and 12 after the addition of 100 µg/mL iron (holo-transferrin) and/or 1 ng/mL TNF-alpha to the calcification medium. The IL-24 protein levels were enhanced by iron and TNF-alpha stimulation, and the increased gene expression level was maintained during the cell culture period. These experiments used one cell lines of HASMCs.