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. 2017 Nov 23;7:16153. doi: 10.1038/s41598-017-16543-7

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© The Author(s) 2017

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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Egr1 leads to inguinal subcutaneous white adipose tissue browning in postnatal and 4 month-old mice. (A–D) Sections of SC-WAT of 1-month-old (A,B) and 4-month-old (C,D) Egr1 ^+/+ and Egr1 ^−/− mice were immuno-stained with UCP1 antibody. Nuclei were visualized with hematoxilin (A,C) or DAPI (B,D). (B,D) upper and lower panels are Dapi/Phase and UCP1/DAPI views of the same field. Scale bars: (A,C) lower magnification 100 µm, higher magnification 50 µm; (B,D) 25 µm. (E) White and beige adipocyte number was counted in arbitrary unit areas of transverse sections of SC-WAT of 1-month-old Egr1^+/+ (N = 10) and Egr1 ^−/− (N = 11) mice and 4-month-old Egr1 ^+/+ (N = 13) and Egr1 ^−/− (N = 14) mice. Graphs show means of counts generated from 10 to 14 sections for each sample ± standard deviations. Asterisks indicate the p-values obtained using the Mann-Whitney test, comparing beige or white adipocyte number between mutant and control mice **P < 0.01, ****P < 0.0001. (F) RT-qPCR analysis of expression levels for beige adipocyte differentiation marker Ucp1 in SC-WAT of 2-week-old and 4-month-old Egr1 ^−/− mice compared to Egr1 ^+/+ mice. Graphs show means ± standard deviations of 5 samples from 2-week-old Egr1 ^+/+ and Egr1 ^−/− mice, 6 samples from 4-month-old wild-type mice and 5 samples from 4-month-old Egr1 ^−/− mice. The Ucp1 mRNA levels of control (Egr1 ^+/+) SC-WAT were normalized to 1. The relative mRNA levels were calculated using the 2^−ΔΔCt method. The p-values were obtained using the Mann-Withney test. Asterisks indicate the p-values *p < 0.05, **p < 0.01. (G) Cell number in SC-WAT in Egr1 ^+/+ and Egr1 ^−/− mice. Number of nuclei (DAPI-positive cells) was counted in arbitrary unit areas of transverse sections of SC-WAT of 1 month-old Egr1 ^+/+ (N = 10) and Egr1 ^−/− (N = 11) mice and 4 month-old Egr1 ^+/+ (N = 13) and Egr1 ^−/− (N = 11) mice. Graphs show means of 10 to 13 sections for each sample ± standard deviations. Asterisks indicate the p-values obtained using the Mann-Whitney test, comparing beige or white adipocyte number between mutant and control mice **P < 0.01, **** P < 0.0001.