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. 2017 Jul 18;7:5756. doi: 10.1038/s41598-017-05810-2

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© The Author(s) 2017

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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Effect of age on the expression of cardiac myosin heavy chains in Hfe ^+/+ and Hfe ^−/− mice. (A) Cardiac myosin expression in 1-month, 5-month and 12-month old Hfe ^+/+ and Hfe ^−/− mice (n = 3–5 per group) was evaluated by fluorescence microscopy. Frozen heart tissues were sectioned using cryostat (10 µm thickness) and stained with bispecific myosin antibody polymer, which was conjugated to anti-dithiopropionic acid-rhodamine isothiocyanate. Sections stained without bispecific polymer were used as background control. (B) Representative immunoblots are shown for α- and β-myosin heavy chains in heart tissues of 1-month, 5-month and 12-month old Hfe ^+/+ and Hfe ^−/− mice (n = 4–6 per group). The bar graph represents the relative expression in the protein level of α- and β-myosin heavy chains, normalized to that of actin. Data are presented as the means ± SEM. *p < 0.05 between Hfe ^−/− and Hfe ^+/+ mice assessed by the two-way ANOVA, followed by the Tukey’s post-hoc analysis.