GSM7886868: LysM-Cre IL10R fl/fl #1; Mus musculus; RNA-Seq
Murine livers were harvested after PBS perfusion and gallbladder removal, cut into small pieces and digested in HBSS (with Ca2+ and Mg2+) containing 10 U/ml DNase and 1 mg/ml Collagenase, in a shaking incubator at 37°C for 25 minutes. After digestion, the livers were smashed and washed using PBS (1% FBS) through a cell strainer to a single cell resolution, and the pellet was collected after centrifugation at 400 g for 8 minutes. Immune cells were then enriched from the pellet by Percoll gradient centrifugation (GE Healthcare, Chicago, IL). Immune cells were then FACS sorted, with myeloid cells being gated as CD45+ CD3- CD11c- CD11b+. RNA was isolated from sorted cells using Trizol (Invitrogen). libraries were constructed using polyA enrichment