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. 2017 Dec 13;8:2115. doi: 10.1038/s41467-017-02162-3

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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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Fig. 4 — SYK deficiency is incompatible with brown fat formation and activation. Genotypes of offspring at weaning age from AdipoQCre Syk^flox/flox mice mated with Syk^flox/flox mice (a). Weight of complete BAT depots from 4 week old mice (n = 5 each genotype) on a chow diet (b). c RNA expression in subcutaneous white adipose tissue scWAT (AdipoQCre Syk^flox/flox, n = 5; Syk^flox/flox, n = 4) and representative immunoblots (d) of BAT (n = 3) from 7–8 week old mice on a chow diet housed at 20 °C. Body weights (e) and glucose tolerance test (f) of AdipoQCre Syk^flox/flox mice or Syk^flox/flox mice on a high fat diet (HFD) in red (AdipoQCre Syk^flox/flox, n = 17; Syk^flox/flox n = 18,) and chow diet (chow) in black (AdipoQCre Syk^flox/flox, n = 15; Syk^flox/flox, n = 21). Asterisks shown are multiple hypothesis corrected p values < 0.05 for an unpaired two-tailed t test of weights (e) or blood glucose (f) per time point. PCR amplification (g) of the deleted Syk^flox allele (top), Syk^flox genotype (middle) and the AdipoQCre allele (bottom) in brown pre-adipocytes (d0) and differentiated brown adipocytes (d4) derived from BAT of the indicated genotype and heterozygous germline deleted mice. h Southern blot of BamHI digested genomic DNA of BAT from 6 weeks old mice with the indicated genotypes. i Oxygen consumption (upper panel) and motor activity (lower panel) before and after intraperitoneal (i.p.) injection of CL316,243 with (n = 13, female mice) or without SYK-i4 (PRT 062607) (n = 13, female mice). j Core body temperature after acute cold exposure with or without i.p. injected SYK inhibitor PRT 062607 (n = 5). k Immunoblotting of BAT from 10 week old CreERT2 Syk^flox/flox mice and Syk^flox/flox controls (n = 4, male mice) 16 days after last tamoxifen treatment. l Oxygen consumption measured (VO₂) during the sleep cycle was compared with VO₂ measured for 12 h post injection with CL316,243 subcutaneous (s.c.) of the mice shown in k (n = 4). For i–j, l P values were calculated with the two-way ANOVA test. For a, P values were determined using a two-tailed chi-square test, (b) data are shown as mean ± SEM, asterisks denote statistical significance (p < 0.05) determined by two-tailed t test and for c, asterisks denote statistical significance (p < 0.05) determined by two-tailed t test