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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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Fig. 1 — a, Phosphoproteomics in livers as per plan in cartoon. b, Heat map and hierarchical clustering of phosphosites across groups indicated in a. c, Phosphoproteome-wide comparisons via z score normalization of phosphosites in green cluster. Grey dots represent individual phosphosites. Blue diamonds represent group means. ***P < 0.001, non-parametric ANOVA (Kruskal–Wallis statistic 797.3, P < 0.0001) followed by Dunn’s multiple comparisons test. d, iGPS prediction of upstream individual kinases that respond to lipids across groups indicated and identified in the green cluster in b. e–g, Pairwise comparisons between indicated groups (fasted versus basal (e), corn oil versus fasted (f), and BODIPY FL C₁₆ versus fasted (g)), showing upregulated or downregulated kinase networks. For a–g, n = 4 mice. For d, darker colour intensity reflects higher kinase score. h, Immunoblots (IB) and quantification for indicated proteins in livers of 2–10-month-old male and female mice that were fed or fasted for indicated durations. N values for number of mice analysed at each timepoint for individual phosphoproteins are indicated in parentheses. P-P70^Thr389/P70: 0 h (n = 27), 3 h (n = 21), 8 h (n = 5), 14 h (n = 16) and 20 h (n = 14); P-S6^Ser235/236/S6: 0 h (n = 26), 3 h (n = 20), 8 h (n = 5), 14 h (n = 16) and 20 h (n = 14); P-AKT^Ser473/AKT: 0 h (n = 26), 3 h (n = 21), 8 h (n = 5), 14 h (n = 16) and 20 h (n = 15); P-SGK1^Thr256/SGK1: 0 h (n = 11), 3 h (n = 9), 8 h (n = 4), 14 h (n = 9) and 20 h (n = 12); P-NDRG1^Thr346/NDRG1: 0 h (n = 17), 3 h (n = 14), 8 h (n = 5), 14 h (n = 12) and 20 h (n = 13); and P-AKT^Thr308/AKT: 0 h (n = 27), 3 h (n = 20), 8 h (n = 5), 14 h (n = 16) and 20 h (n = 15). Ponceau is loading control. Individual replicates and means are shown. *P < 0.05 and **P < 0.01, one-way ANOVA followed by Tukey’s multiple comparisons test (h). Please refer to Supplementary Table 10 statistical summary, and Supplementary Tables 1 and 2. Source numerical data are available in Source Data Extended Data Table 1, and unprocessed blots are available in the Source Data for this figure.

Source data