Single cell Transcriptomic analysis of CD8+ T cells from bone marrow chimeras reconstituted with CD137 wild-type or knock out bone marrow, and implanted with B16K1 melanoma.

We report the single cell RNA sequencing of mouse CD8+ T cells in bone marrow chimeras reconsitituted with equal amounts of CD137-/-tomato+ and CD137-/-tomato- bone marrow implanted with B16K1 melanoma. CD8 T cells from the spleen or the tumor infiltrate are compared. (CD137-/-tomato+) and (CD137+/+tomato-) mixed BM chimera were injected s.c. with B16K1 melanoma cells. tomato+ and tomato- spleen and tumor infiltrating CD8+ T cells were isolated using FACSaria™ Fusion (BD biosciences). Single-cell libraries (5′ gene expression) were generated from CD8 cell fractions from 7 mice per group hashed together, using the Chromium Controller Instrument and Chromium Next GEM Single Cell 5’ Library & Gel Bead Kit v1.1 according to the manufacturer’s protocol (10× Genomics). Single-cell library size and quality were confirmed on the Tapestation 4200 (Agilent) and pooled to reach at least 20,000 read pairs for 5’ Gene Expression Library; 10,000 read pairs of Protein Library. Libraries were sequenced twice on a NextSeq 550 (Illumina) in pair-end sequencing 26 bp (read1) × 91 bp (read2) and a single index 8 bp in length.