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. 2018 Jun 8;8:8780. doi: 10.1038/s41598-018-26646-4

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Smad 7 is a target gene of miR-410-5p. (a) Complementarity between the 3′UTR of Smad 7 gene and miR-410-5p. For constructing of Smad 7-3′UTR (Mut) reporter, the putative miR-410-5p binding site was replaced with the sequence as shown below (Mut). Mutated seeds were marked in red, Watson-Crick base pairing with a straight line and U-G wobbles with a dotted line. (b) The putative target sites of Smad 7 3′-UTR, which are marked in red, are conserved in mammals. (c) Luciferase analysis of a reporter vector harboring the 3′-UTR of Smad 7 (WT and Mut) in H9c2 cells transfected with negative control (NC) or miR-410-5p for 36 h. These results are means ± SD of three experiments. **P < 0.01 relative to NC. (d) Western blot analysis in H9c2 cells transfected with NC or miR-410-5p for 36 h. Representative Western blots are shown. (e) Quantification of the ratio of proteins to β-actin. These results are the means ± SD of four experiments. *P < 0.05; **P < 0.01 relative to NC.