Bulk RNA-seq analysis of iWAT cells from Prrx1-ERT; tdTomato mice
One day post Tamoxifen injection, inguinal WATs were washed with PBS, minced, and digested with type II collagenase (2 mg/ml) in KRBB containing 10 mM HEPES (pH 7.4) and 3% BSA for 1 hr at 37C. The digested tissues were filtered through 70-m and 40-m cell strainers (BD Biosciences) and then centrifuged at 500 g for 5 min. The cell pellet was resuspended in PBS for FACS sorting.  5 x 10^5 Tomato+ cells were sorted using S3e Cell Sorter equipped with a 561 nm wavelength laser and performed bulk RNA sequencing.
PET_iWAT_2