Transcriptional differences between intratumoral CD8+ T cells in young and aged tumors at single cell resolution
Single-cell RNA sequencing of  intratumoral CD8+ T cells (and CD45+ cells)  from young and aged mice Overall design: Live tumor-infiltrating CD8+ T cells (or OT-1 CD8+ T cells) and/or intratumoral CD45+ cells were sorted from tumors on day 15 after tumor inoculation by BD FACSSymphony S6 Sorter (BD). Sorted cells were pooled into groups (five biological replicates per group) with independent mice being labeled with TotalSeq-B Hashtags (Biolegend). Cells were counted and loaded onto the Chromium Controller (10X Genomics) for a target recovery of 10,000 single cells per run. Samples were then processed using the 10X Single Cell 3'v3.1 kit per the manufacturer's protocol. Sequencing was done on an Illumina NovaSeq S2 or NextSeq 1000/2000 sequencer using 100-cycle kits with paired-end reads.