GSM7431953: IL-18R-CD103-GFP- cells; Mus musculus; RNA-Seq
Lungs were transcardically perfused through the right ventricle with PBS to remove blood from the organ. Each lobe of the lung was separated and digested in 1 mg/ml collagenase type III (Worthington), 200 g/ml DNase I (Roche), 0.4 units/ml Dispase (Gibco) in RPMI1640 medium plus 2% FCS with gentleMACSTM Dissociator (Miltenyi Biotec) using the mouse lung digestion protocol setting. The digested lung tissue was then spun down and the mononuclear cells were isolated on a 40%-80% Percoll gradient. Lymphocytes were recovered from the interface and were washed twice with PBS. Live CD45+lin-CD90.2+IL-7R+ were purified using flow cytometry. Library was prepared using 10X Genomics protocols using the Chromium Next GEM Single Cell 3' Reagent Kits v3.1 (Dual Index)