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. 2023 Jun 29;25(7):989–1003. doi: 10.1038/s41556-023-01163-3

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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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Extended Data Fig. 1 — (a) Direct BODIPY fluorescence in liver slices of 8 mo-old C57BL/6 male mice subjected to 30 min gavage with vehicle or 10 mg/kg of BODIPY FL C₁₆ after 14–16 h fasting (n = 5 mice). (b) Dendrogram of hierarchical clustering analysis (Euclidean distance) between experimental groups based on phosphorylation status of 863 phosphosites. (c) Prediction for kinases that putatively target the phosphosites identified within the green cluster in main Fig. 1b. Darker color intensity reflects higher kinase score. (d) Pairwise comparisons showing upregulated (red) or downregulated (blue) kinase networks in mice refed high fat diet (HFD) for 30 min after 14–16 h fasting. For b-d (n = 4 mice). (e) Representative IB, and (f) quantification for the indicated proteins in livers of 2–8 mo-old C57BL/6 male or female mice kept fasted for 14–16 h or treated with corn oil or BODIPY FL C₁₆ or refed a HFD for 30 min after fasting. N values for number of mice analyzed for individual proteins and conditions are indicated in parentheses. RAPTOR and P-AMPK^Thr172/AMPK: n = 4 per group; RICTOR: fasted (n = 8), corn oil (n = 8), BODIPY FL C₁₆ (n = 4), refed (n = 4); P-P70^Thr389/P70: fasted (n = 23), corn oil (n = 22), BODIPY FL C₁₆ (n = 8), refed (n = 4); P-S6^Ser235/236/S6 fasted (n = 23), corn oil (n = 24), BODIPY FL C₁₆ (n = 9), refed (n = 4); P-AKT^Ser473/AKT: fasted (n = 17), corn oil (n = 13), BODIPY FL C₁₆ (n = 9), refed (n = 4); P-AKT^Thr308/AKT: fasted (n = 4), corn oil (n = 3), BODIPY FL C₁₆ (n = 4), refed (n = 4). Ponceau is the loading control. (g) Circulating free fatty acids (FFA) in 2–7 mo-old male mice fasted for: 0 h (n = 20 mice), 3 h (n = 9 mice), 14 h (n = 3 mice) or 20 h (n = 15 mice). Individual replicates and mean values are shown. *P < 0.05, ***P < 0.001 and ****P < 0.0001, One-way ANOVA followed by Šídák’s multiple comparisons test (f); One-way ANOVA followed by Tukey’s multiple comparisons test (g). Please refer to Supplementary Table 10_statistical summary, and Supplementary Tables 1, 2. Source numerical data are available in SourceData_Table 1, and unprocessed blots are available in Source Data Extended Data Fig. 1.

Source data