GSM6081051: Ctrl_1; Mus musculus; RNA-Seq
Ear skin was first digested in 5 mg/ml Collagenase II in PBS supplemented with 0.2 mg/ml DNaseI and 0.2% FBS, followed by filtering through 50 µm filters (CellTricks). Fc-receptors were blocked with rat anti-mouse CD16/32 antibody (eBioscience), followed by staining using Podoplanin-APC and CD31-Pe-Cy7 antibodies. Dump channel included erythrocytes and immune cells (labeled using CD45-eF450, CD11b-eF450 and Ter119-eF450) and dead cells (SYTOX Blue dead stain (Life Technology)). The sorting into 384-well plates was performed with a 100 µm nozzle on BD FACS AriaIII CellSorter (BD BioScience Flow Cytometry System equipped with four lasers: 405, 488, 561, 633 nm). Smart-Seq2 library preparation according to Picelli S et al. Full-length RNA-seq from single cells using Smart-seq2. Nat. Protoc. 2014;9(1):171–181.