title: BAT_TN-HFD_4
organism: Mus musculus
description: Protocols: All experiments were approved by the Animal Ethics Committee of the North Stockholm region. The experiments were performed on mice exposed to different physiological conditions (described below). All groups had free access to water and food and 12:12-h light-dark cycle regime. The analysis of samples from different physiological conditions was performed in parallel, as indicated in the corresponding experiments. Standard conditions. C57BL/6 male mice were bought from Charles River and were kept at 21 °C until 8 weeks of age. The samples analyzed in this study were from the wild-type mice (C57BL/6) described and metabolically characterized in 25. Before the start of the experiment, the mice were housed at 22-24 °C. At the start of the experiment, 12-week-old male mice were single-caged and transferred to 30 °C (thermoneutrality). The mice had access only to high-fat diet (45 % calories from fat, Research Diets D12451). The mice were kept at thermoneutrality for at least 25 weeks. Tissues were harvested and snap frozen. Total RNA was extracted from frozen adipose tissue (left lobe/pad) with TRI Reagent® (Sigma-Aldrich) according to the manufacturer's protocol, and RNA concentrations were measured on a Nanodrop nd-1000 spectrophotometer (Thermo-Scientific, Wilmington, DE). TapeStation was applied to validate the RNA integrity. Sequencing libraries were prepared from 100 ng of total RNA (RIN 7.5-9.4) using the TruSeq Stranded mRNA Library Prep Kit (Illumina). Quality and concentration of NGS libraries were verified using the Agilent 2200 TapeStation.