GSM5651473: Fibp_3_KO; Mus musculus; RNA-Seq
CD8+ T cells were isolated from the single-cell suspension of splenocytes of Pmel-1 TCR transgenic mice using the EasySep mouse CD8+ T cell isolation kit (STEMCELL, Cat# 19853) by magnetic negative selection. If cells were not used directly, the freshly isolated splenocytes would be frozen in cryopreservation medium (Gibco, Cat# 2176664). CD8+ T cells isolated from newly-thawed splenocytes using the same method were rested in the medium overnight before stimulation. Isolated CD8+ T cells were stimulated with plate-bound anti-mouse CD3 (TONBO, 40-0032-U100, clone 17A2) at 10 ug/mL and anti-mouse CD28 (TONBO, 40-0281-U100, clone 37.51) at 5 ug/mL for 48 hours before expanded with recombinant mouse IL-2 (Biolegend, 575406) at 100 IU/mL. RNA libraries were prepared for sequencing using standard Illumina TruSeq Stranded mRNA-seq library protocol