GSM8988561: Lung, Pdgfrb, mutant, scRNAsq; Mus musculus; RNA-Seq
Lungs from young adult mice were freshly collected after perfusion and kept in cold 1X Leibovitz media until dissociation. All the lung lobes except for the accesory lobe were separated and dissociated in 1X Lebovitz media with Collagenase type I (2mg/ml), DNAse I  (0.5 mg/ml) and Elastase (2mg/ml) for 45  minutes at 37˚C. Then, the lung cells were filtered, red blood cells lysated and lung cells were incubated  with conjugated antibodies for each lineage for 30 min. All the samples were stained with CD45-PE/Cy7 (BioLegend, 103114), ECAD-PE (BioLegend, 147304), and ICAM2-A647 (Invitrogen, A15452) antibodies (1:250 dilutions for all antibodies) for 30 min Then washed and stain with the viability dye Sytox blue. Cells were filtered with a 35 mm cell strainer into a 5 mL glass tube again and sorted by a BD FACSAria  with a 70 mm nozzle. FACS-purified lung cells were processed through the Chromium Single Cell Gene Expression Solution Platform (10x Genomics) using the Chromium Single Cell 3' Library and Gel Bead Kit in accordance with the manufacturer's user guide. The libraries were sequenced on an Illumina NextSeq500 using a 26X124 sequencing run format with 8 bp index.