GSM8077397: UHE-HL-g003; Mus musculus; RNA-Seq
Lungs were digested in DMEM/F12 containing 4 U/ml Dispase and 10 µg/ml DNaseI at 37°C for 45 min. Digested tissues were filtered through a 100 µm cell strainer. Erythrocytes were lysed using ACK buffer. An anti-mouse CD16/32 dimer antibody was employed to inhibit non-specific binding of antibodies to FcγII and FcγIII. The cell suspension was incubated with anti-CD31 MicroBeads to pre-select PECAM-1high cells using MACS LS columns and a magnetic separator. Samples were cryopreserved and thawed according to 10x Genomics guidelines. Cells were washed, filtered, counted, and loaded at a concentration of 700–1200 cells/µl on a 10x Genomics chip. The chip was then run on a Chromium controller to initiate GEM formation. Library preparation was performed, following standard 10x Genomics guidelines