Effect of Grk2 (spleen, rat) or Ets1 (spleen and spinal cord parenchyma, rat; in vitro, human) KOs on gene expression of CD4+ T cells

3' bulk RNAseq was conducted on CD4+ T-MBP rat cells isolated form spleen or spinal cord parenchyma three days after co-transfer of control and KO cells into the animal. Cells were FACS sorted based on their fluorescent protein reporters, BFP for control, and GFP for KOs. For human experiments, CD4+ T cells were isolated from buffy coats, KOed and restimulated in vitro, and 3' bulk RNAseq was performed. For rats, three biological replicates were used from three different animals, for humans, four biological replicates were used from four different donors本研究对共移植对照与敲除（KO）细胞至动物体内三天后分离得到的CD4阳性T-MBP大鼠细胞（CD4+ T-MBP rat cells）开展3'端批量RNA测序（3' bulk RNAseq）：此类细胞从脾脏或脊髓实质中分离，随后依据荧光蛋白标记物通过荧光激活细胞分选术（Fluorescence-Activated Cell Sorting, FACS）完成分选，其中携带蓝色荧光蛋白（Blue Fluorescent Protein, BFP）的为对照组细胞，携带绿色荧光蛋白（Green Fluorescent Protein, GFP）的为敲除组细胞。针对人体实验，研究人员从血沉棕黄层（buffy coat）中分离CD4阳性T细胞，对其进行基因敲除处理后开展体外再刺激，随后同样完成3'端批量RNA测序。大鼠样本设置3个生物学重复，分别取自3只不同的实验动物；人体样本设置4个生物学重复，分别取自4名不同的供体。