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. 2018 Feb 21;8:3377. doi: 10.1038/s41598-017-17540-6

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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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Inhibition of lysosomal or endosomal function augments osteogenic VSMC transformation. Confluent rat aortic VSMCs were cultured in Pi + Ca²⁺ (3.5 mM phosphate/3 mM calcium) calcification medium with or without 10 μM of lysosomal inhibitor chloroquine or 40 μM of endosome inhibitor dynasore for 7 days, and the fresh calcification culture medium was changed every other day. (A–C) qPCR results demonstrate effect of chloroquine on osteoblast markers RUNX2 and BMP2, and VSMC contractile marker TAGLN in Pi + Ca²⁺-stimulated VSMCs. (D–F) qPCR results showed an enhanced effect of dynasore on Pi + Ca²⁺-induced osteogenic transformation. The data represent 6 determinations and were replicated two times. Values are presented as mean ± SD. *P < 0.05.