GSM8230798: CD8_TILs_WildType4; Mus musculus; RNA-Seq
CD4+ and CD8+ tumor infiltrating T lynphocytes were isolated from B16F10 subcutaneuos tumors from WT and Sirt2KO mice (n = 4 per group). Total RNAs were extracted using the RNeasy Micro Kit (Qiagen; cat. #74004) according to the manufacturer's instructions. Extracted RNAs were quantitated with the Qubit Fluorometer (ThermoFisher Scientific, Waltham, MA) and screened for quality on the Agilent TapeStation 4200 using the high-sensitivity RNA ScreenTape (Agilent Technologies, Santa Clara, CA). The samples were then processed for RNA-sequencing using the Takara SMARTer Stranded Total RNA-seq Kit v2 Pico Input Mammalian kit (Takara Bio USA Inc., San Jose, CA). Briefly, one ng of RNA was used to generate cDNA and a strand-specific ribosomal RNA-depleted library following the manufacturer's protocol. Quality control steps were performed, including TapeStation size assessment and quantification using the Kapa Library Quantification Kit (Roche, Wilmington, MA). The final libraries were then normalized, denatured, and sequenced on the Illumina NextSeq 2000 sequencer with the P3-200 cycle reagent kit to generate at least 60 million 100-base read pairs per sample (Illumina, Inc., San Diego, CA).