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. 2018 Mar 1;9:890. doi: 10.1038/s41467-018-03196-x

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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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Fig. 1 — Mural Pparg overexpression drives healthy visceral WAT expansion in obesity. a Pdgfrb^rtTA; TRE-Pparg2 (Mural-Pparg^TG) mice are generated by breeding the Pdgfrb ^rtTA transgenic mice to animals expressing Pparg2 under the control of the tet-response element (TRE- Pparg2). Littermates carrying only Pdgfrb^rtTA or TRE-Pparg2 alleles were used as the control animals for Mural-Pparg^TG. Pdgfrb^rtTA; TRE-Cre; Rosa26R^mT/mG; TRE-Pparg2 (MuralChaser-Pparg^TG) mice are generated by breeding the “MuralChaser” (Pdgfrb^rtTA; TRE-Cre; Rosa26R^mT/mG) mice to animals carrying TRE-Pparg2 transgene. MuralChaser mice were used as control animals for MuralChaser-Pparg^TG. b Control and Mural-Pparg^TG mice were fed a standard chow diet until 6 weeks of age before being switched to dox-containing high-fat diet (HFD). Body weights were measured weekly following the onset of HFD feeding. Control-HFD, n = 8; Mural-Pparg^TG-HFD, n = 9. Data points represent mean + s.e.m. c Fat mass and lean mass (normalized to body weight) of control and Mural-Pparg^TG mice after 16 weeks of dox-HFD feeding. Control-HFD, n = 8; Mural-Pparg^TG -HFD, n = 9. Bars represent mean + s.e.m. d Fat pad weight (normalized to body weight) of control and Mural-Pparg^TG mice after 16 weeks of dox-HFD feeding. * denotes p < 0.05 from Student’s t-test. Control-HFD, n = 8; Mural-Pparg^TG-HFD, n = 9. Bars represent mean + s.e.m. e–h Representative immunofluorescence staining of Perilipin (green) and Mac-2 (red) in e, f gonadal and g, h perirenal WAT paraffin sections obtained from control and Mural-Pparg^TG mice after 16 weeks of dox-HFD feeding. Scale bar, 200 μm. i Number of crown-like structures (Mac-2 positive) in the indicated depots from control and Mural-Pparg^TG mice after 16 weeks of dox-HFD feeding. * denotes p < 0.05 from Welch’s t-test. n = 24 randomly chosen ×10 magnification fields from six individual animals. Bars represent mean + s.e.m. j Average adipocyte size in indicated fat depots from control and Mural-Pparg^TG mice after 16 weeks of dox-HFD feeding. * denotes p < 0.05 from Student’s t-test. n = 6 per genotype. Bars represent mean + s.e.m. k–r Representative ×63 magnification confocal immunofluorescence images of k–n gonadal and o–r perirenal WAT sections from MuralChaser and MuralChaser-Pparg^TG mice after 16 weeks dox-diet feeding. Sections were stained with anti-GFP (green) and anti-Perilipin (red) antibodies and counterstained with DAPI (blue; nuclei). * indicates GFP-labeled perilipin-positive cells. Scale bar, 50 μm. s–t Percentage of perilipin-positive adipocytes expressing GFP in s gonadal and t perirenal WAT from MuralChaser and MuralChaser-Pparg^TG mice maintained on dox-diets for 16 weeks. Two-way ANOVA, *p < 0.05; n = 6 individual depots per group. Bars represent mean + s.e.m. u, v Relative mRNA levels of indicated genes in u gonadal and v perirenal WAT obtained from control and Mural-Pparg^TG mice after 16 weeks of dox-HFD feeding. * denotes p < 0.05 from Welch’s t-test. n = 6 per genotype. Bars represent mean + s.e.m.