Single-Cell Sequencing Data of CD45+ HET Cells
Animal procedures were conducted in compliance with Institutional Animal Care and Use Committee (IACUC) of University of Pennsylvania. WT (C3-T+ Elf5+/+) and HET (C3-T+ Elf5+/) FVB mice used in this study. Tumor tissues from age matched mice were minced and digested by collagenase from Clostridium histolyticum (300 units ml1; SigmaAldrich; catalogue number C2674) and hyaluronidase (100 units ml1; SigmaAldrich; catalogue number H3884) in base medium DMEM/F-12 from Invitrogen (catalogue number 11330032) supplemented with epidermal growth factor (Novoprotein; catalogue number CO29), insulin (Life Sciences; catalogue number 51500056), (cholera toxin, SigmaAldrich; C8052) and hydrocortisone (SigmaAldrich; catalogue number h0888). Post-whole tumor tissue digestion, tumor cells were brought to single cells using 0.25% trypsin EDTA and Dispase/DNAse solution (4 mg DNAse (SigmaAldrich) and 200 mg ispase (Invitrogen) in 40 ml PBS) sequentially. 5% heat-inactivated FCS in PBS was used in between to stop the enzymatic reactions. Red blood cells were removed using 0.64% sterile ammonium chloride solution for 5 min at 37 C. Tumor cells were further passed through a 0.45-m strainer to obtain single cells. Selective enrichment of NK cells for bulk RNA seq and CD45+/- cells for single cell RNA seq were performed by sorting in Aria B cell sorter.