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. 2023 Dec 14;25(1):378–403. doi: 10.1038/s44319-023-00012-6

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© The Author(s) 2023

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/. Creative Commons Public Domain Dedication waiver http://creativecommons.org/publicdomain/zero/1.0/ applies to the data associated with this article, unless otherwise stated in a credit line to the data, but does not extend to the graphical or creative elements of illustrations, charts, or figures. This waiver removes legal barriers to the re-use and mining of research data. According to standard scholarly practice, it is recommended to provide appropriate citation and attribution whenever technically possible.

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(A) Database analysis of 2750 genes associated with NAFLD and their rankings. (B) A Venn diagram showing the overlap between the NAFLD genes and mitochondrial proteins that are highly expressed in liver tissue. Eighty proteins, including NME4, were identified. (C) Line chart showing the gene expression in liver compare with mean of all tissue identified in (B). (D) H&E staining and Oil Red O staining of liver sections of the mice fed a HFD for the indicated times. Scale bars, 50 μm. (E) The steatosis area of H&E staining (D) was quantified by Image-Pro Plus (IPP). Biological replicates, n = 3. (F) Oil Red O-positive area of (D) was quantified by Image-Pro Plus (IPP). Biological replicates, in 2 w n > 5; in 6 w n = 5; in 18 w, n = 11. (G) Immunohistochemical staining of liver sections from the mice fed a HFD for the indicated times. Scale bars, 50 μm. (H) Nme4-positive area was quantified by Image-Pro Plus (IPP). Biological replicates, in 2 w n = 5; in 6 w n = 3; in 18 w, n = 5. (I) Western blots probed with antibodies against Nme4 and Actin in the livers of mice fed a normal diet or HFD for 18 weeks and 24 weeks. (J) Relative mRNA levels of Nme4 in the livers of the mice fed a normal diet or HFD for 6 weeks and 18 weeks were measured by RT‒qPCR. The mRNA level was detected by qPCR and normalized to β-actin. Biological replicates, n = 3. (K) The correlation between the Nme4-positive area and the steatosis area was analyzed using Pearson’s correlation, P values was detected by two-tailed Student’s t test (paired). (L,M) Primary hepatocytes were treated with different doses of PO (PA: OA = 0.4 mM : 0.8 mM) for different times. Blots probed with antibodies against Nme4 and Actin are shown. Data information: (E,F,H,J) data are presented as mean ± SEM. *P values ≤ 0.05, **P values ≤ 0.01, ***P values ≤ 0.001 (Student’s t test, unpaired). HFD high-fat diet, w week, h hour, PA palmitic acid, OA oleic acid. Source data are available online for this figure.