title: GSM8036812: Bbs8 KO iWAT, APC P2, n1; Mus musculus; RNA-Seq
library_construction_protocol: Progenitor cell isolation from the stromal vascular fraction (SVF) was performed as previously described (Sieckmann et al., 2022).Tissues were minced and digested with collagenase II and DNAse I at 37°C for 30 min. The digestion was quenched by adding AT buffer (0.5% BSA in PBS). The dissociated cells were passed through a 100-μm filter (Corning). The stromal vascular fraction was resuspended in red blood cell lysis buffer (BioLegend). Isolated cells were then further subjected to antibody staining. Flow sorting of cell was done by gating to isolate single cells distinct from debris, doublets, and dead cells. The cells were further gated to exclude the lineage (CD31, CD45, TER119) and to enrich for CD29 and Sca1 to isolate pre-adipocytes. Subpopulations were sorted by CD26 and CD55 for the P1 population, CD54 for P2 subpopulation, and CD142 for P3. After sorting, cells were stored in Trizol. Total RNA was extracted using the miRNeasy Micro kit (Qiagen) according to manufacturer's protocol. Smart‐seq02 (Picelli et al. 2014)