GSM7518422: Adipose cDC1s, LFD_DC1; Mus musculus; RNA-Seq
Adipose tissues were physically dissociated using  scissors and incubated in a shaking incubator (225rpm) at 37°C for 45 min in adipose digest solution (2mg/ml type II collagenase in HBSS +MgCl+CaCl, 5% fetal calf serum, 1% L-glutamine, 1% penicillin-streptomycin, and 50ug/mL DNase1). The resulting dissociated tissue was passed through 100μm strainers, and adipocytes were removed from the supernatant by centrifugation. Red blood cells were lysed using ACK lysis buffer and the resulting stromal vascular fraction pellets were processed to single cell suspensions for FACS. cDC1s were sorted to to > 95% purity as via as follows: cDC1: (CD45+Lin–(CD3-TCRβ-CD19-Ly6G-NK1.1-)CD88-HLA-DR+CD11c+CD11b–XCR1+ RNA libraries were prepared for sequencing using standard Illumina protocols.