GSM7978054: LLC WT + 20 GY IR, Gene Expression Library, scRNAseq; Mus musculus; RNA-Seq
Freshly collected lungs of tumor-bearing mice were minced and dissociated with 1 mg/mL collagenase IV and 200 μg/mL DNaseI at 37°C for 45 minutes. Red blood cells were removed with lysis buffer. Samples were then filtered through a 70 μm cell strainer and washed twice with sorting buffer (PBS supplemented with 2% FBS and 0.5 mM EDTA). The cells were re-suspended in staining buffer and were blocked with anti-FcR and stained with live/dead dye and extracellular antibodies. Live CD45+ cells were sorted with a BD FACSAria Fusion 5-18 and constantly kept on ice. Cells were re-blocked with anti-FcR and barcoded using TotalSeq™-B0301, B0302, B0303 and B0304 hashtag antibodies Library construction was performed according to the manufacturer's instruction (10x genomics)