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The ETS2 plasmid, encoding the full-length human ETS2 gene, was obtained from Addgene (28128). Truncated plasmids were constructed using Gibson assembly Master Mix (E2621; New England Biolabs). MicroRNA-223 (miR-223) reporter plasmid was obtained by subcloning the miR-223 promoter into pGL3-Basic vector. Luciferase assays were performed using the luciferase assay system (E1980; Promega). The Adeno-X Adenoviral System 3 kit (632267; Clontech) was used to generate an adenoviral vector expressing ETS2 (AdETS2) or mutated ETS2 (AdETS2 T72A and AdETS2 T72E).
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Circulation 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.This article is made available via the PMC Open Access Subset for unrestricted re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the COVID-19 pandemic or until permissions are revoked in writing. Upon expiration of these permissions, PMC is granted a perpetual license to make this article available via PMC and Europe PMC, consistent with existing copyright protections.
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