Genotypical comparison of T<sub>reg</sub> and T<sub>conv</sub> cells isolated from brown adipose tissue (BAT) and spleen tissue (SPL) in cold- and warm-conditioned animals generated with an Illumina Mouse Expression Array.
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Abstract
<p>(A) Gene expression profiles comparing T<sub>reg</sub> (top) or T<sub>conv</sub> (bottom) cell populations between spleen and adipose tissue samples isolated from warm-conditioned animals (left) or between cells isolated from cold vs warm-conditioned animals (right). Numbers indicate genes either up- or downregulated more than 2-fold (cut-off: dotted line), with the number of significantly different (p<0.05) genes shown in brackets with an asterisk. (B) Volcano plot comparing gene expression and significance values between T<sub>reg</sub> and T<sub>conv</sub> genes isolated from BAT in warm-conditioned animals. Key up- or downregulated genes in T<sub>reg</sub> cells are annotated (Foxp3, Il10, Cxcl1/2, Tcf7, Ifng) and serve as quality control to the published consensus T<sub>reg</sub>-cell signature. (C) Hierarchical clustering of the top-30 upregulated genes and the top-10 downregulated genes in warm-conditioned brown adipose tissue T<sub>reg</sub> cells versus spleen T<sub>reg</sub> cells. (D) Comparison of BAT-T<sub>reg</sub>-specific genes with visceral adipose tissue (VAT)-specific genes. We first determined 430 genes to upregulated in BAT warm-conditioned T<sub>reg</sub> cells, with 222 genes being significantly altered (p<0.05). We then overlaid BAT T<sub>reg</sub>-upregulated genes with VAT T<sub>reg</sub> tissue specific expression gene data. 181 genes were matched between both microarary chips, with 169 genes also upregulated in VAT, and only 12 genes specific for BAT (left). The corresponding analysis of the 516 genes upregulated in cold BAT T<sub>reg</sub> cells versus warm spleen T<sub>reg</sub> cells revealed 194 genes to be significantly upregulated. 158 could be matched to VAT T<sub>reg</sub>-specific genes, of which 148 were VAT-specific, whereas only 10 were specific for BAT. P-values indicate the significance of overrepresentation of BAT T<sub>reg</sub>-specific genes in the VAT T<sub>reg</sub> signature. (E) Comparison of VAT-T<sub>reg</sub> specific genes on BAT warm (left) or BAT cold (right) gene signatures. Of 1839 genes specifically overexpressed in VAT T<sub>reg</sub> cells, 1059 were statistically significantly (p<0.05) upregulated. Of these 1059 genes, 829 were also detectable in the BAT T<sub>reg</sub> microarray. When comparing the VAT T<sub>reg</sub> signature to warm BAT T<sub>reg</sub> cells, 660 genes were overrepresented in VAT, whereas cold BAT T<sub>reg</sub> cells show 685 genes to be overrepresented in VAT.</p
