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. 2014 May 20;3(3):e000796. doi: 10.1161/JAHA.114.000796

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© 2014 The Authors. Published on behalf of the American Heart Association, Inc., by Wiley Blackwell.

This is an open access article under the terms of the Creative Commons Attribution‐NonCommercial License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.

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Figure 8. — Binding of Shox2a to the Nkx2.5 regulatory elements depends on its phosphorylation. ChIP assays reveal 2 Shox2a‐binding elements, site‐1 (A) and site‐2 (B), within the 3.3‐kb Nkx2.5 promoter region. (C) ChIP assays using mouse embryonic hearts confirm binding of Shox2a to the site‐2 in vivo. D, Electrophoretic mobility shift assay (EMSA) reveals specific binding of the site‐2 sequence by nuclear extracts from cells expressing HA‐Shox2a, but not HA‐Shox2a‐DM. Red arrows point to the shift bands. (E) Supershift assay with anti‐HA antibodies demonstrates specific DNA with HA‐Shox2a. Green arrow points to the supershifted band. ChIP indicates chromatin immunoprecipitation.