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. 2019 Apr 16;9:6165. doi: 10.1038/s41598-019-42127-8

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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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Brown adipocyte-specific CXCR4-deficient mice were generated. Expression of mRNA extracted from each tissue or primary brown adipocyte was evaluated using qRT-PCR and normalised to 36B4 gene. Serum CXCL12 concentration was measured using the Quantikine ELISA Mouse CXCL12/SDF-1α. Values are shown as mean ± SD. Student’s t-test was used to compare with a difference. *P < 0.05. n.s.: not a significant difference. Black and open bar or circle represent control and the knockout mice group, respectively. WT: wild-type, CXCR4^flox/−mice; KO: knockout, CXCR4^flox/floxmice (A) knockout mouse in using the Cre-loxP system. Exon 2 of CXCR4 was deleted in the BAT expressed UCP-1 gene. (B) CXCR4 mRNA expression in primary brown adipocyte from CXCR4^flox/flox compared with CXCR4^flox/− (n = 3). (C) Expression of CXCR4 gene in the brown adipose tissue (BAT), Liver, Intraperitoneal white adipose tissue (WAT), and subcutaneous WAT (n = 7). (D) UCP-1, PGC-1α, and Cytochrome C1 mRNA expression in the BAT of CXCR4^flox/− and CXCR4^flox/flox mice (n = 6). (E) UCP-1 protein level in the BAT of CXCR4^flox/− and CXCR4^flox/flox mice (n = 4–6). (F) Intraperitoneal glucose tolerance test in the mice with normal chow diet. (G) Insulin tolerance test in the mice with normal chow diet. (H) Serum CXCL12 concentration in mouse at nine weeks old (n = 6–5) and 30 weeks old (n = 9–7). (I) Change of food intake in normal chow diet. (J) Change of food intake in the high-fat chow diet. (K) Change of body weight in the mouse with normal chow diet.