GSM8606019: tumor_ctrl_Tcells_1; Mus musculus; RNA-Seq
Murine pancreatic cancer cells mPDAC were injected in syngenic immunocompetent FVB/n mice. Mice were treated with anti-TNFR2 mAb or untreated. After 18 days, mice were euthanized, and tumors were processed with Collagenase IV and DNase I. Cells were stained with DAPI, CD45, CD4, CD8, CD11c, Ly6G, Ly6C, CD19, NK1.1, F4/80 and T cells were sorted on a FACSAria III (BD) by gating on live cells, CD45+, lineage (CD11c, Ly6G, Ly6C, CD19, NK1.1, F4/80) negative cells, CD4+ and CD8+ cells. CD4 and CD8 isolated T cells were mixed. Sorted T cells samples were used for scRNA-seq analysis. Indexed libraries were tested for quality, equimolarly pooled and sequenced on an Illumina HiSeq2500 using paired-end 26 × 98bp as sequencing mode. By using a full Rapid flow cell, coverage was around 100M reads per sample corresponding to 100,000 reads per cell.