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. 2019 Dec 23;9:19696. doi: 10.1038/s41598-019-56051-4

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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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WSB/EiJ mice show no increase of inflammatory responses to HFD in the hypothalamic arcuate nucleus (ARC) in contrast with C57BL/6J mice, but comparable responses in the hypothalamic paraventricular nucleus (PVN). Immuno-histochemical analysis of microglia cells in the hypothalamic nuclei of twelve old C57BL/6J and WSB/EiJ male mice either maintained under control feeding (CTRL), or challenged with HFD for the last 3 days of the period (3D) or during the whole 8-week experiment duration (8WK). (A) Representative confocal images of ARC immune-labelled with Iba1 (in green), a microglia marker, and DAPI-stained nuclei (in blue), in coronal sections of C57BL/6J (top panel) and WSB/EiJ (bottom panel) mice. Regions of interest (ROI) are delimited in white. Microglia pointed by arrowhead were magnified in insets at the top left of the main image; (B) Quantitative analysis of microglia area (top; soma + ramifications, in µm²) and density (bottom; number of microglia per µm² of ROI) obtained from confocal images showed in A (N = 10–24 sections/group). Graphs show median values with range in scatter dot plot. Significant differences are indicated by different letters to account for between strain and between diet group differences (P ≤ 0.05; GLM); (C) Representative confocal images of PVN immune-labelled with Iba1 (in green), a microglia marker, and DAPI-stained nuclei (in blue), in coronal sections of C57BL/6J (top panel) and WSB/EiJ (bottom panel) mice. Regions of interest (ROI) are delimited in white. Microglia pointed by arrowheads were magnified in insets at the bottom left of the main image; (D) Quantitative analysis of microglia area (top; soma + ramifications, in µm²) and density (bottom; number of microglia per µm² of ROI) obtained from confocal images showed in C (N = 10–24 sections/group). Graphs show median values with range in scatter dot plot. Significant differences are indicated by different letters to account for between strain and between diet group differences (P ≤ 0.05; GLM); Scale bars in main images: 100 µm; scale bars in insets = 10 µm.