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. 2021 Apr 20;12:2330. doi: 10.1038/s41467-021-22431-6

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© The Author(s) 2021

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. 4 — a–f Distributions of fatty acids and phospholipids in the hypothalamus in regular chew diet- (RCD-) or high-fat diet- (HFD-) fed mice for 8 weeks. a, d Representative results of IMS on hypothalamic arachidonic acid (AA) (a) and PI (18:1/20:4) (d) from RCD-fed mice (left) or HFD-fed mice (right). Scale bar: 500 μm. b, c Relative intensities of fatty acids in the VMH (b) or ARC (c) of RCD- (n = 7) or HFD-fed mice (n = 5; two-tailed t test for each molecule, ARC: p = 0.0190 in AA(20:4), p = 0.0104 in OA(18:1), p = 0.0387 in SA(18:0), RCD vs HFD). e, f Relative intensities of phospholipids in the VMH (e) or ARC (f) of RCD- (n = 7) or HFD-fed (n = 5) mice (two-tailed t test for each molecule, for VMH: p = 0.0196 in PI (18:0/20:4), p < 0.0001 for PI (18:1/20:4), p = 0.0362 for PE (18:0/20:4), p = 0.0080 for PE (p18:0/20:4), p = 0.0307 for PS (18:0/22:6), for ARC: p = 0.0325 for PI (18:0/20:4), p = 0.0061 for PI (18:1/20:4), p = 0.0282 for PE (18:0/20:4), p = 0.0192 for PE (p18:0/20:4), and p = 0.0347 for PS (18:0/22:6)). g, h Enzymatic activity of hypothalamic cPLA2 (g) and sPLA (h) in RCD- (n = 5) or HFD-fed (n = 5) mice (two-tailed t test, p = 0.0264 in g, RCD vs HFD). i–k Relative amount of total-cPLA2 (t-cPLA2) and phosphorylated-cPLA2 (p-cPLA2) between RCD and HFD-fed mice. i Representative photos of western blotting. j, k Quantification of p-cPLA2 (j) and t-cPLA2 (k) between RCD (n = 4) and HFD (n = 3) fed mice (two-tailed t test, p = 0.0338 in j, RCD vs HFD). l Relative amounts of prostaglandins in the hypothalamus after 8 weeks in HFD-fed mice (n = 3) compared with those of RCD-fed mice (n = 3). Major prostaglandins were underlined. m–s Bar graphs showing COX-mediated production of AA (m), PGD2 (n), PGF2α (o), PGE2 (p), 11-beta-13,14-dihydro-15-keto-PGF2α (q), 13,14-dihydro-15-keto-PGD2 (r), and 20-hydroxy-PGF2α (s) in 8 weeks of HFD-fed mice (n = 3) compared with RCD-fed mice (n = 3; two-tailed t test, p = 0.0256 in n, p = 0.0339 in o, p = 0.0013 in p, p = 0.0354 in q, p = 0.0260 in r, p = 0.0250 in s, RCD vs HFD). All data represent the mean ± SEM; *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.