GSM8386413: Eosinophil_specific_GPR43_KO; Mus musculus; RNA-Seq
To isolate asthmatic lung cells, lungs were chopped into small fragments and enzymatically digested with Collagenase D and DNase I at 37 °C for 30 minutes. The enzyme reaction was stopped by adding EDTA. After vigorous resuspension, cells were filtered through 100 µm strainers, and red blood cells were lysed with ACK lysis buffer. The dead cells and FSC(hi)SSC(hi) cells were removed by fluorescence-activated cell sorting (FACS). To synthesize scRNA-seq libraries, cells were suspended in PBS containing 0.04% BSA and loaded into Chromium-controller (10X Genomics) scRNA-seq libraries were generated using the Chromium Next GEM Single Cell 3ʹ Reagent Kits v 3.1 (10X Genomics) according to the manufacturer's protocol