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. 2017 Jul 24;7:6312. doi: 10.1038/s41598-017-06395-6

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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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Immunolocalization of myospryn and minispryn in isolated cardiomyocytes. Enzymatically isolated guinea pig ventricular cardiomyocytes were fixed and processed for confocal immunofluorescence microscopy using the indicated antibodies. (a–c) At low magnification myospryn (a) immunoreactivity is visible in discontinuous striations that run perpendicular to the long axis of the cardiomyocyte partially overlapping the nucleus (arrow in a). (d–f) Higher magnification images show that myospryn immunoreactivity is present in distinct punctae that co-localise with ryanodine receptors (e) in the SR (f and inset). (g–i) Anti-minispryn (g) immunoreactivity also appears in discontinuous striations running the length of the cardiomyocyte and is also present in the nucleus (arrow in g). (j–l) In common with myospryn, higher magnification images show that minispryn partially co-localises with ryanodine receptors (k) in the SR (l and inset). (m–o) Myospryn immunoreactivity overlaps the Z-line labelled with an anti-alpha-actinin antibody (m, n, o). (p–r) Similarly, minispryn immunoreactivity also partially overlaps alpha-actinin at the Z-line (p, q, r). (s–x) Cardiomyocytes labelled with either anti-SERCA2 (s, t, u) or anti-sarcalumenin (v, w, x) antibodies and myospryn antibodies demonstrate that myospryn immunoreactivity (arrowheads in s) appears to be restricted to the j-SR rather than the l-SR (arrows in t).