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. 2018 Nov 9;8:16594. doi: 10.1038/s41598-018-34808-7

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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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Physiological expression of APP and APLP2 in heart muscle cells of the left ventricle. (A) With an antibody directed against the N-terminus of APP there was a mild cytoplasmic staining in all cardiomyocytes of a left ventricle sample from case No. 15 exhibiting longitudinal-skewed cut muscle fibers. One BD lesion was seen (arrow) (B) In these cells lipofuscin particles (black arrowheads) were also stained as seen at the higher magnification level. The BD-inclusion (arrow) was morphologically different from the lipofuscin particles because it was bigger and showed a homogenous mass of strongly APP-positive material in the mildly APP expressing muscle cells. The nuclei of the cardiomyocytes were not stained and serve as intrinsic negative control for this staining (red arrowheads). (C) The Aη-D-epitope of APP was also expressed in all cardiomyocytes, here shown in a sample of the left ventricle myocardium of case No. 15. The muscle fibers were cut transversally. (D) At higher magnification level lipofuscin particles were also stained with the antibody against the Aη-D-Epitope (black arrowheads). BD-lesions were not seen with this antibody. The nuclei of the cardiomyocytes were not stained and served as intrinsic negative control for this staining (red arrowheads). (E,F) The Aβ epitope of APP was only faintly stained in cardiomyocytes of the left ventricle myocardium of case No. 58 using an antibody raised against Aβ_1–17. No BD-lesions could be seen. Lipofuscin was not stained as well. The nuclei of the cardiomyocytes were not stained and served as intrinsic negative control for this staining (red arrowheads). The muscle fibers were cut transversally. (G) APLP2 was expressed in the cytoplasm of all cardiomyocytes of the left ventricular myocardium of case No. 1 exhibiting muscle fibers in longitudinal-skewed orientation. (H) At the higher magnification level lipofuscin particles (black arrowheads) were stained with this antibody as well. BD-lesions could not be detected with this antibody. The nuclei of the cardiomyocytes were not stained and served as intrinsic negative control for this staining (red arrowheads). (I,J) Negative controls by omitting the primary antibodies for biotinylated anti-mouse IgG (I) and anti-rabbit IgG secondary antibodies (J) were performed on left ventricular myocardium of case No. 44. Positive controls are shown in Suppl. Fig. 1.