GSM3103162: SUB HFD 2; Mus musculus; RNA-Seq
Adipose tissues were excised from mice and immediately minced in a collagenase solution comprising 0.2% collagenase (C6885, Sigma) and 2% BSA dissolved in Hanks' balanced salt solution (HBSS, Gibco). Minced tissues were transferred to a 50ml tube and incubated at 37ºC for 20min (for EPI and ING) or 40min (for BAT) at 500rpm. 10ml complete DMEM was subsequently added. Cell resuspensions were filtered through 100μm strainers, spun at 2000rpm for 5min, and washed once with PBS. The floating adipocyte layer and the pelleted SVF fraction were collected separately. Total RNA was extracted using Qiagen kit. 1ug of total RNA was used for each RNA-Seq library preparation according to the manufacturer's instructions (New England Biolab), and sequencing was done on HiSeq 2000 (Illumina).