GSM6560313: VBCS-581_WT5; Mus musculus; RNA-Seq
Total RNA was extracted with a combination of TRIzol and PureLink RNA mini kit (Invitrogen). For eWAT: Libraries were prepared by the Van Andel Genomics Core from 500 ng of total RNA using the KAPA Stranded mRNA-Seq Kit (v4.17) (Kapa Biosystems, Wilmington, MA USA). RNA was sheared to 300-400 bp. Prior to PCR amplification, cDNA fragments were ligated to IDT for Illumina TruSeq UD Indexed adapters (Illumina Inc, San Diego CA, USA). For macrophages: Libraries were prepared by the Van Andel Genomics Core from 1-5 ng of total RNA. Total RNA material was converted to dsDNA using the SMART-Seq v4 Ultra Low Input RNA Kit for Sequencing, v. 091817 (Takara Bio USA, Mountain View, CA, USA ). cDNA was sheared to 200–500 bp using a Covaris E220 Evolution ultrasonicator (Covaris Inc., Woburn, MA USA) and Illumina compatible sequencing libraries were generated using the Kapa HyperPrep kit, v6.17 (Kapa Biosystems, Wilmington, MA USA). Prior to PCR amplification, cDNA fragments were ligated to Bioo Scientific NEXTflex dual adapters (Bioo Scientific, Austin, TX, USA).