RNA-seq Analysis of Soluble Tim-3 Treatment on CD8+ T Cells. Mus musculus

OT-I CTLs were pretreated with sTim-3 protein, followed by stimulation with anti-CD3 and anti-CD28 antibodies. Then cells were harvested and snap-frozen immediately in TRIzol, awaiting RNA-seq analysis. Following the manufacturer protocol, NEBNextUltra RNA Library Prep Kit for Illumina was used to generate the sequencing libraries before adding the index codes to attribute sequences to each sample.

将OT-I型细胞毒性T淋巴细胞（OT-I CTLs）用可溶性T细胞免疫球蛋白黏蛋白3（soluble Tim-3, sTim-3）蛋白预处理，随后以抗CD3抗体与抗CD28抗体进行刺激。收集细胞后立即使用TRIzol试剂进行速冻保存，以待开展RNA测序（RNA-seq）分析。按照试剂盒制造商的操作规范，使用适配Illumina平台的NEBNextUltra RNA文库制备试剂盒构建测序文库，随后添加索引序列以将测序序列归属至对应样本。