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. 2016 Mar 9;7:22. doi: 10.3389/fendo.2016.00022

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Copyright © 2016 Janssen, Zuidwijk, Muller, van Mil, Dirkx, Oudejans, Paulus and Simonides.

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) or licensor 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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Dio3 is targeted by miR-214. A dual-luciferase construct containing the complete Dio3 3′UTR downstream of the Renilla luciferase gene was used to analyze the interaction between miR-214 and the 3′UTR of Dio3. Renilla luciferase activity (RLuc) is expressed relative to firefly luciferase activity (FLuc), correcting for differences in transfection efficiency. Four independent transfections were performed in duplicate. No-miR: control without co-transfection; ctrl-miR: + negative control/scrambled miR-1; P214: + Pre-miR-214; PA214: + Pre-miR-214 and Anti-miR-214. Values are means ± SEM, n = 4, normalized to the RLuc/Fluc ratio in the no-miR group, *p < 0.05.