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. 2022 Mar 29;12:5341. doi: 10.1038/s41598-022-09428-x

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© The Author(s) 2022

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 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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Identification of MDBs in vitro. (A) Representative images of HE staining and immunofluorescence staining. Eosinophilic bodies and CK8/18 aggregates were observed in glucotoxicity, lipotoxicity, and glucolipotoxicity groups. However, p62-positive bodies were identified only in the glucotoxicity and glucolipotoxicity groups. Red arrows indicate MDBs. Scale bar, 100 μm in normal views. Scale bar, 50 μm in enlarged views. HE: Hematoxylin–eosin, CK: Cytokeratin. (B) p62 protein quantification in PHH/HSC sheets by ELISA. p62 levels were significantly higher in the lipotoxicity and glucolipotoxicity groups compared to control and glucotoxicity groups. n = 4, 3 independent experiments, **p < 0.01, *p < 0.05. (C) Representative images of TEM. Red arrows indicate MDBs. MDBs were observed in the glucotoxicity and glucolipotoxicity groups. Scale bar, 10 μm. N: nuclear. (D) Representative images of TEM. Giant mitochondria was observed in the glucolipotoxicity group. Scale bar, 2 μm. M: mitochondria, GM: giant mitochondria with crystalline inclusion.