GSM7372336: CD45, anti-PD1; Mus musculus; RNA-Seq
Murine tumors were harvested and minced into smaller pieces using a sharp blade before incubation with 1mg/mL Collagenase IV (Roche) for 30 minutes at 37C with frequent agitation. After incubation, 10mL of fresh culture media was added before filtration through a 70μm filter. Following centrifugation, cell pellets were resuspended in 3mL of red blood cell lysis solution (BioLegend) on ice for 5 min followed by the addition of 10mL complete media. Single cell suspensions were then washed with PBS, and stained with Zombie-NIR LIVE/DEAD stain and CD45 antibody for 30 min at 4oC. Live/CD45+ leukocytes were sorted by BD Biosciences Aria II cell sorter with 100-mm nozzle. Library was performed according to the manufacter's instructions (single cell 3' v3 protocol, 10x Genomics).