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Relative mRNA level MGP/18S

*. 6. MGP. EV. Pi + + + + TNF - - + + EV MGP EV MGP. 30. 4. Relative mRNA level MGP/18S. *. 20. 2. Calcium/protein ( μ g/mg). 10. 0. EV. MGP. 0. Pi. Pi+TNF.

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Relative mRNA level MGP/18S

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  1. * 6 MGP EV Pi + + + + TNF - - + + EV MGP EV MGP 30 4 Relative mRNA level MGP/18S * 20 2 Calcium/protein (μg/mg) 10 0 EV MGP 0 Pi Pi+TNF B C A Supplemental figure 1. Role of MGP overexpression in Pi/TNF-induced calcification. Identification the overexpression of MGP (A) by retroviral vector in HASMCs with real-time RT-PCR; EV, empty vector. (B) Calcium content assay and (C) alizarin red staining showed the role of MGP overexpression on calcification. Pi + TNFα Ctrl Pi TNF No virus Ad-GFP Ad-IκBα von Kossa ANKH p-p65 Supplymental figure 2. ANKH expression in aortic rings was inhibited by TNF. Human renal artery tissue was cultured in DMEM medium with Pi (3.0 mM) and/or TNF (10 ng/ml) for 2 weeks. For adenovirus-IκBα (Ad-IκBα) or –GFP (Ad-GFP) group, tissue was infected with corresponding adenovirus for 24 hours before treatment. Von kossa staining showed calcium deposition, and immunohistochemical staining detected ANKH and p-p65 (s536) levels. One of the representative images from each group was shown (×100).

  2. Supplemental Table 1. Serum biochemical measurements of rats Supplemental Table 2. Primers for Quantitative PCR.

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