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. 2018 Dec 3;8:17547. doi: 10.1038/s41598-018-35890-7

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© The Author(s) 2018

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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Microgliosis and markers of neuroinflammation in the brain of western diet-fed Sirt3^−/− mice. Wild type and Sirt3^−/− mice were fed on standard (SD) and western diet (WD) for four months. (a) Brain samples were subjected to immunofluorescent staining of Iba1 with FITC (green) for microglia. The nuclei were stained with DAPI (blue). Representative images, captured at 400X magnification in a Leica confocal microscope, are presented. (b) The mRNA of IL-1β was determined by real-time PCR in an ABI Prism 7700 sequence detector (Applied Biosystems, Foster City, CA). (c) Immunohistochemical analysis of IL-1β was performed with the mouse brain samples. Representative images are presented. (d) Western blot analysis of IkBα was performed with the mouse brain samples. The blots were reprobed with β actin antibody. (e) The band intensity of IkBα was quantitated by densitometric scanning and corrected for the levels of β actin. The values are mean ± SE of six observations. *P < 0.01; **P < 0.001 vs wild type mice on standard diet.