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Total RNA was extracted with Trizol (Thermo Fisher Scientific). Subsequently, cDNA was synthesized with a PrimeScript RT Master Mix (Takara, Japan). The mRNA expression levels were quantified by qPCR on CFX connect real-time PCR system (Bio-rad), using THUNDERBIRD SYBR qPCR Mix (Toyobo, Japan) and were normalized to the expression of GAPDH. The 2−ΔΔCt method was used to calculate the relative expression levels of mRNA. Sequences of the primers were listed in the Data Supplement.
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Arteriosclerosis, Thrombosis, and Vascular Biology is published on behalf of the American Heart Association, Inc., by Wolters Kluwer Health, Inc. This is an open access article under the terms of the Creative Commons Attribution Non-Commercial-NoDerivs License, which permits use, distribution, and reproduction in any medium, provided that the original work is properly cited, the use is noncommercial, and no modifications or adaptations are made.
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