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. 2023 Oct 12;14(10):673. doi: 10.1038/s41419-023-06170-8

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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 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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Fig. 1 — A Hepatic levels of AF6 in healthy patients (n = 20) and in patients with simple steatosis (n = 11) and non-alcoholic steatohepatitis (NASH) (n = 10) in GSE89632. B mRNA levels of Af6 in mice maintained on a high-fat and high-cholesterol (HFHC) diet for 16 weeks. C Gene set enrichment analysis (GSEA) and D heatmap of necroptosis in HFHC model. E Immunoblot of AF6, RIPK1, MLKL, MLKL oligomer, p-MLKL, USP21, and Hsp90 in liver after maintenance on HFHC diet. Densitometric analysis was used to quantify band intensities; the protein levels of AF6 and p-MLKL were normalized to that of the housekeeping protein Hsp90 in the respective lanes. Correlation between AF6 and p-MLKL levels was assessed using Graphpad Prism 8.0. Data are expressed as mean ± SEM. Pairwise comparisons between groups were conducted using two-tailed non-paired Student’s t tests. ns, not significant (p ≥ 0.05); *p < 0.05; **p < 0.01; ***p < 0.001.