GSM8908260: Male_Ads_Ctnnb1flfl, rep 1; Mus musculus; RNA-Seq
Epididymal WAT (eWAT) and posterior subcutaneous WAT (psWAT) depots were excised from Ctnnb1fl/fl and Ctnnb1AdKO mice as described [51]. eWAT and psWAT depots were minced with scissors and then digested for 20 and 30 min, respectively, at 37°C in 500 U/ml collagenase type I (Worthington Biochemical) in Krebs-Ringer-HEPES (KRH; pH 7.4) buffer containing 1 g/l glucose and 500 nM adenosine with shaking (100 rpm). Cell suspensions were filtered through 300 μm cell strainers and then centrifuged at 80 x g for 8 min. Buoyant adipocytes were separated from the pelleted SVCs. Fractions were washed three times with KRH buffer. Total RNA was isolated and purified from adipocytes or SVCs using RNA STAT-60 (AMSBIO) according to the manufacturer's instructions. After DNAse treatment, samples were subjected to quality control, library preparation, and strand-specific mRNA sequencing (Beijing Genomics Institute, China).