RNA sequencing (RNA-seq) of memory CD8 T cells and CD8+/− MR1tet+ cells was performed as described previously (31 (link)). Briefly, total RNA was purified using an miRNeasy Micro Kit (QIAGEN) and quantified by quantitative PCR, as described previously (32 (link)). Purified total RNA (1–5 ng) was amplified following the Smart-Seq2 protocol (33 (link)). mRNA was captured using poly-dT oligonucleotides and directly reverse transcribed into full-length cDNA (amplified by PCR for 16 cycles). cDNA was purified using AMPure XP beads (Beckman Coulter). From this step, 1 ng of cDNA was used to prepare a standard Nextera XT sequencing library (Nextera XT DNA Sample Preparation Kit and Index Kit; Illumina). Whole-transcriptome amplification and sequencing library preparations were performed in a 96-well format to reduce assay-to-assay variability. Quality-control steps were included to determine total RNA quality and quantity, the optimal number of PCR preamplification cycles, and fragment size selection. Samples that failed quality control were eliminated from further downstream steps. Barcoded Illumina sequencing libraries (Nextera; Illumina) were generated using the automated platform (Biomek FXp). Libraries were sequenced on an HiSeq 2500 Illumina platform to obtain 50-bp single-end reads (TruSeq Rapid Kit; Illumina).