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. 2019 May 6;9:6909. doi: 10.1038/s41598-019-43335-y

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© The Author(s) 2019

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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Reduced lipophagy in Plin2^−/− hearts. (A) Immunblot analysis of LC3BI, LC3BII, p62 and Lamp-1 in lysates from Plin2^+/+ and Plin2^−/− hearts after O/N fasting. Full-length blots are presented in Supplementary Fig. S10. (B) Quantification of immunoblot analysis in A (n = 4). (C) Immunblot analysis of LC3BI, LC3BII and p62 in cardiomyocytes, treated with or without 25 µM chloroquine, isolated from Plin2^+/+ and Plin2^−/− mice. Full-length blots are presented in Supplementary Fig. S11. (D) Quantification of immunoblot analysis in C (n = 4). (E) Representative confocal microscopy images of Plin2^+/+ and Plin2^−/− cardiomyocytes, shown as a max stack containing 10 z-stack images compressed into one single image, and a higher magnification z-stack image. Lamp-1 (lysosomes) is shown in green and ORO (LDs) is shown in red, nuclei in blue, and co-localization of Lamp-1 and ORO as as yellow (x630 magnification, scale bar 10 µm, inset image scale bar 1 µm). (F,G) Quantification of co-localization between lysosomes and lipid droplets. (F) Quantification of Manders coefficient tM1 (fraction of ORO in co-localization with Lamp-1) in cytosol and plasma membrane, and (G) Manders coefficient tM2 (fraction of Lamp-1 in co-localization with ORO) in cytosol and plasma membrane in cardiomyocytes isolated from Plin2^+/+ and Plin2^−/− mice, (n = 4–5). Data are presented as mean ± SEM, *p < 0.05 vs. Plin2^+/+.