Assay for Transposase-Accessible Chromatin using sequencing of spleen memory CD4+ T cells

We performed a ATAC seq analysis of 2 biological independent splenic samples from pooled mice sorted for lcmv-specific memory CD4 T cells and 2 technical replicates from one mouse sorted for naive CD4 T cells . Overall design: Cells negative for dump and positive for CD4, CD44 and gp66 and either Ly6C+ PSGL-1+ (Th1) Ly6C-PSGL-1- (TFH) or dump- CD4+ CD44- CD62L+ for naive CD4 T cells were sorted for each replicate, and used for ATAC Seq

本研究针对2份生物学独立的脾脏样本开展了ATAC测序（ATAC-seq）分析，样本来自混合小鼠，分选得到淋巴细胞性脉络丛脑膜炎病毒（LCMV）特异性记忆CD4 T细胞；同时针对1只小鼠的样本完成2份技术重复，分选得到初始CD4 T细胞。整体实验设计如下：各重复均分选以下两类细胞群体：其一为dump阴性且表达CD4、CD44及gp66的细胞，进一步分为Ly6C+PSGL-1+（T辅助细胞1型，Th1）与Ly6C-PSGL-1-（滤泡辅助T细胞，TFH）两个亚群；其二为初始CD4 T细胞，即dump阴性、CD4+、CD44-且CD62L+的细胞。分选得到的细胞均用于后续ATAC测序分析。