title: RNA-seq of Homo sapiens: exhausted CD8+ T cell
design: CD8+T cells were isolated using Human CD8+ T Cell Isolation Kit (Miltenyi Biotec) according to the manufacturers procedure. Purified CD8+T cells were then stimulated with anti-CD3/anti-CD28-coated microbeads (Dynabeads M-450 Epoxy, Invitrogen; bead to cell ratio of 1:1) in the presence of 10U/ml recombinant human IL-2 (PeproTech) in T cell medium (RPMI 1640 medium supplemented with 10% FBS, 50M -mercapteoethanol, 1M HEPES, 1X non-essential amino acids (NEAA), 100mM sodium pyruvate, and 1% penicillin-streptomycin). Surviving cells were passaged every 48 hours with fresh microbeads and IL-2 until day 8 to generate Tex cells. Alive Tex cells were sorted through FACs with Influx (BD Biosciences). Their RNA was extracted using RNeasy Mini Kit (Qiagen) and submitted for paired-end RNA sequencing. Reads were mapped to UCSC human genome GRCh38/hg38 using STAR (version 2.6.1) and gene counts were quantified by featureCounts (version 1.6.4). Control group: exhausted CD8+ T cells, replicate 3