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. 2020 Jun 22;10:10052. doi: 10.1038/s41598-020-66903-z

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© The Author(s) 2020

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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Total phosphorylation levels for MyBP-C, MLC-1, desmin & tropomyosin increased with age in males, while only MyBP-C and troponin-T increased with age in females. (A) MyBP-C phosphorylation increased moderately with age in males and dramatically with age in females; MyBP-C phosphorylation also was greater in middle-aged males compared to females. (B) MLC-1 phosphorylation increased markedly with age in males but not females; values were higher in older males than age-matched females. (C) Desmin phosphorylation increased with age in males only; values were significantly higher in older males compared to older females. (D) Tropomyosin phosphorylation increased with age in males only. (E) Phosphorylation of troponin-T increased with age in females only and was significantly higher in older females compared to older males. (F) Phosphorylation of troponin-I was not affected by either age or sex. Values denote the mean ± SEM in each case. Data were analyzed with two-way ANOVA (age and sex were main factors; the post-hoc test was Holm-Sidak). The * denotes p < 0.05. Samples were hearts from 5 middle-aged male mice, 6 older males, 5 middle-aged females and 5 older females, respectively. Actin was used as a loading control; it was not affected by either age or sex (not shown). In all cases data were normalized to actin. Myosin-binding protein C (MyBP-C), myosin light chain-1 (MLC-1).