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. 2019 Feb 8;10:662. doi: 10.1038/s41467-019-08607-1

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© The Author(s) 2019

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. 5 — Consequences of growth hormone receptor (GHR) ablation in leptin receptor (LepR)-expressing cells or the entire brain. a–c A high percentage of LepR neurons (red) in the arcuate nucleus (ARH; t₍₅₎ = 28.42, P < 0.0001), lateral hypothalamic area (LHA; t₍₅₎ = 6.777, P = 0.0011), dorsomedial nucleus (DMH; t₍₅₎ = 10.51, P = 0.0001) and ventral premammillary nucleus (PMv; t₍₅₎ = 14.6, P < 0.0001) is responsive to porcine growth hormone (GH) (20 µg/g body weight (b.w.)) in control mice (phosphorylation of signal transducer and activator of transcription 5 (pSTAT5), green), whereas very few pSTAT5 is observed in tdTomato cells of LepR GHR KO mice (n = 3–4; unpaired t-test). Yellow represents double-labeled cells. Scale Bar = 50 µm. d GHR mRNA expression in the hypothalamus of control and brain GHR KO mice (t₍₁₂₎ = 30.02, P < 0.0001, n = 6–8; unpaired t-test). e Body weight changes in control, LepR GHR KO and brain GHR KO mice (main effect of time [F_(10, 1036) = 370.9, P < 0.0001], main effect of GHR ablation [F_(2, 1036) = 76.28, P < 0.0001] and interaction [F_(20, 1036) = 0.4301, P = 0.9867]; two-way analysis of variance (ANOVA); n = 14–20; *P < 0.05, LepR GHR KO vs. control mice; ^#P < 0.05, brain GHR KO vs. control mice; ^†P < 0.05, LepR GHR KO vs. brain GHR KO mice). f Body length (F_(2, 42) = 16.29, P < 0.0001, n = 13–17). g Body adiposity (F_(2, 34) = 9.815, P = 0.0004, n = 11–14). h Serum leptin concentration (F_(2, 30) = 4.477, P = 0.0199, n = 10–12/group). i Lean body mass (F_(2, 38) = 13.17, P < 0.0001, n = 7–22/group). j Hypothalamic GHRH mRNA expression (F_(2, 21) = 13.43, P = 0.0001, n = 7–9) of 6-month-old male mice. One-way ANOVA and the Newman–Keuls test were used when the data of control, LepR GHR KO and brain GHR KO mice were compared. All results were expressed as mean ± s.e.m.