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. 2021 Oct 20;12:692431. doi: 10.3389/fphar.2021.692431

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Copyright © 2021 He, Qu, Kwan Law, Qiu, Han, Ricardo de Seabra Rodrigues Dias, Liu, Zhang, Wu, Wu, Fai Mok, Cheng, He and Wai Wong.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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AGEs induce calcification in HASMCs. (A) Dose-dependent study of AGEs-induced calcification in HASMCs at AGEs concentrations from 0 to 100 mg/L. Von Kossa staining (black) in HASMCs after AGEs treatment for 2 weeks. scale bar = 100 µm (×10). (B) Immunofluorescence staining of calcification-related proteins with 24 h’ AGEs treatment at indicated concentrations. The green GFP signal represents the calcified protein OPN. The blue DAPI signal represents the nuclear region, scale bar, 25 µm. (C) Western blot analysis of the expression of RAGE and other calcification-related proteins (β-catenin, OPG, OPN, MMP7, BMP2, Runx2) with 24 h’ treatment. Data are presented as the fold change compared with the control group. Each point represents the mean ± SD (n = 3/group); ^* p < 0.05, ^** p < 0.01, ^*** p < 0.001.