Single cell sequencing (Gene expression, TCR sequencing, and antigen specificity of CD8 T cells from T3 sarcoma
T3 tumor-bearing mice were treated on day twelve post-tumor transplant. Two and four days later, single-cell suspensions were prepared from TILs. Total CD8+ T cells were enriched using a CD8+ T cell positive selection kit (Miltenyi). Tumors from individual mice were labeled with one of the following antibodies before pooling in order to identify cells from individual tumors in downstream analysis: 1) TCR (TotalSeq-C0120 anti-mouse TCR  chain antibody, Cat# 109259, BioLegend), 2) CD90.2 (TotalSeq-C0075 anti-mouse CD90.2 (Thy1.2) antibody, Cat#, 105353, BioLegend), and 3) CD45 (TotalSeq-C0096 anti-mouse CD45 antibody, Cat# 103169, BioLegend). 10x Genomics libraries were generated using 10x 5'v2 Single Cell RNAseq, V(D)J enrichment, feature barcoding, and Barcode Enabled Antigen Mapping kits.