RNA-Seq Analysis of LCMV-Specific CD8+ T Cells

Cross-linking was performed using 5 million sorted LCMV-Specific CD8⁺ T cells 12 dpi according to a published protocol (Guo et al., 2014 (link)). RNA immunoprecipitation was performed with 10 µg pan-AGO antibody (MABE56; Millipore) and Ago2 antibody (015–22031; Wako Chemical), and quantitative RT-PCR was performed following a published protocol (Guo et al., 2015 (link)). RT reaction was performed using the Invitrogen Superscript III cDNA synthesis kit according to the manufacturer’s protocol. Sybr-based quantitative PCR was performed with the following primers: set-1 forward, 5′-GCAGTTCAGCCAAGACAGAG-3′: set-1 reverse, 5′-TGTAGTGATACATACGTAGAGTGCAA-3′; set-2 forward, 5′-CTGCAAGTGCCATCCTTGTA-3′; set-2 reverse, 5′-TGACCTAAAATTAAATGAATGCAAA-3′; set-3 forward, 5′-TTTAAAAGGTGCCCGCACTA-3′; set-3 reverse, 5′-TGCATCACTTCAAGTTCCTTCA-3′; set-4 forward, 5′-GGCAGCAGTTCCTTAGTTTACA-3′; set-4 reverse, 5′-GCCCAAATGATCAACGTCAT-3′; set-5 forward, 5′-GGCAGAATCAGTGTTCGTGA-3′; set-5 reverse, 5′-CAACAAACGAATCAACAACTGC-3′; set-6 forward, 5′-CAGTAGAGATGCAGTTGGTTCC-3′; set-6 reverse, 5′-AAAACTGGGGAAAGGGAGAA-3′; set-7 forward, 5′-AGGTTACAGGAGGCTGGATG-3′; set-7 reverse, 5′-TGCTCTGTGAAGGGAATTCTG-3′; set-8 forward, 5′-TTTGGTTCACAGCCGTTTTC-3′; and set-8 reverse, 5′-AAAAGTACGTGTCAGTAAGAAGGGTA-3′.