GSM5005588: KD-C57-Sp; Mus musculus; RNA-Seq
For tumors established by i.v. injected tumor cells, lung tissues were digested by 2 mg/ml collagenase I for 30 minutes and homogenized by pressing through a 70 μm cell strainer to generate single cell suspensions. Cell suspensions of tumor tissues were subjected to a discontinuous Percoll gradient (70%/40%) by centrifugation at 2000 rpm without break at room temperature for 20 minutes. Mononuclear cells at the interface were collected and washed with PBS before further analysis.CD8+ T cells were isolated from the mononuclear cells or spleen using immunomagnetic separation beads according to the manufacturer's instructions. RNA libraries were prepared for sequencing using standard 10x genomics protocols