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. 2018 Sep 13;8:13730. doi: 10.1038/s41598-018-31877-6

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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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AGEs accelerated VSMC calcification through a PDK4-dependent pathway. (A) PDK4 siRNA transfection efficiency was determined by western blotting. *P < 0.05 vs. the indicated treatment. (B) Calcified VSMCs were transfected with PDK4 siRNA or scrambled siRNA for 24 h and then cultured with or without AGE-BSA (200 μg/ml) for another 24 h. RUNX2 expression was determined by western blotting. *P < 0.05 compared with the normal control group. ^#P < 0.05 compared with the AGE-BSA +β-GP group. (C) After transfection, calcified VSMCs were cultured with or without AGE-BSA (200 μg/ml) for 7 days, and ALP activity and calcium deposition were detected. *P < 0.05 compared with the normal control group. ^#P < 0.05 compared with the AGE-BSA +β-GP group. (D) Calcified VSMCs were incubated with or without AGE-BSA (200 μg/ml) and DCA for 21 days. Calcium nodule formation was visualized by Alizarin red S staining.