GSM9512735: Knockout; Mus musculus; RNA-Seq
12 week-old male mice were fed chow diet supplemented with 600 mg/kg doxycycline for 2 weeks and SVF cells were isolated from sWAT as described above. SVF was harvested from sWAT as described above. After centrifugation the cells were resuspended in 2% FBS in PBS containing propidium iodide (PI; 1:400) for 15 min incubation on ice. After washing, the cells were resuspended in 2% FBS in PBS for sorting of viable cells, PI- cells. FACS was conducted with a BD Bioscience FACSAria cytometer at the Flow Cytometry Core Facility at UT Southwestern. Single-cell RNA-seq libraries were prepared using the 10x Genomics Chromium Single Cell 3′ Gene Expression v3.1 (Dual Index) kit according to the manufacturer's protocol and sequenced on an Illumina NextSeq 2000 with paired-end reads (28 bp Read 1, 90 bp Read 2; 10 bp i7 and 10 bp i5).