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. 2020 Apr 10;11(9):661–679. doi: 10.1007/s13238-020-00713-x

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© The Author(s) 2020

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Mitochondria in cardiomyocytes of Bmal1-deficient mice exhibited increased fusion and decreased mitophagy. (A) Significantly enlarged mitochondria were observed in Bmal1-deficient myocardium comparing to those in the wild type group. Arrows indicate mitochondria. (B and C) Mitochondrial size and area-perimeter ratio were carried out for the quantification of mitochondrial fusion level (enlarged and fused mitochondria possess greater area-perimeter ratio). (D) The protein expression level of MFN2 was calculated by western Blotting analysis evaluation (expressed as the ratio of β-ACTIN). (E) Columns in graphs show protein normalized for β-ACTIN. (F) Less mitophagy take place in Bmal1^−/− mice myocardium than in wild type control group. (G) Mitophagy were counted in 12 different transmission electron images of Bmal1^−/− mice myocardium and wild type control group, respectively. (H and I) The protein expression level of LC3A/BI/IIand BNIP3 were assessed by Western Blotting analysis evaluation (expressed as the ratio of LC3A/BII and BNIP3 to β-ACTIN). Data represented the mean ± SD. *P < 0.05, **P < 0.01 and ****P < 0.0001 versus wide type by two-tailed Student’s t test