GSM6402519: hypoxic Vegfr2Y949F A set, sample 2, scRNAseq; Mus musculus; RNA-Seq
Fresh lungs were dissected from mice following 6-week hypoxia exposure, minced and subjected to collagenase/DNase digestion as described above. The dissociated cell suspension was filtered through a 70-μm strainer and then a 40-μm strainer. Thereafter, red blood cells were lysed and removed from the cell suspension and the remaining pulmonary cells were centrifuged at 1,500 rpm for 5 minutes, rinsed with 0.04% BSA/PBS for 3 times, and re-suspended in 0.04% BSA/PBS A 10X Genomics analysis system was used to prepare single-cell whole transcriptome. Single cell capture was achieved by nanodroplet (single-Poisson distribution loading)-based encapsulation of 10X barcoded gel beads with the cells. The resulting gel beads in emulsion (GEMs) containing the captured cells were then retrieved, dissolved and subjected to reverse transcription of polyadenylated mRNAs. Single-cell transcriptomic sequencing libraries were generated from 10X barcoded cDNAs and feature barcode DNAs and sequenced on a NovaSeq System (Illumina) with a 150 bp paired-end run according to the manufacture's instruction