GSM8783895: 2Gyx5, biological replicate 4; Mus musculus; RNA-Seq
Mice were humanely euthanized using CO2. Tumors were excised and minced with sterile razor blades and then enzymatically dissociated with papain (#LS003, Worthington) at 37 degrees Celsius for 45 minutes. Samples were subsequently centrifuged for 5 minutes at 500g, resuspended in RBC lysis buffer (#00-4300-54, eBioscience), incubated for 10 minutes at room temperature, and then resuspended in 5% FBS in PBS. Cell suspensions were serially filtered through 70 mm and 40 mm filters before being resuspended again in 5% FBS in PBS for manual cell counting using a hemacytometer. A total of 10,000 cells were loaded per single-cell RNA sequencing sample onto the 10x platform using the Chromium Single Cell 3' Library & Gel Bead Kit v3.1 on a 10X Chromium controller (10X Genomics) per the manufacturer recommended default protocol and settings.