Single cell RNA-sequencing of Mbd3-null and control lymphoid progenitor cells

To determine the role of Mbd3/NuRD in lymphopoiesis, gene expression in single Mbd3-deleted and control lymphoid progenitor cells was analysed using single cell RNA-seq Overall design: Single Mbd3-deficient and control lymphoid progenitors were isolated from mouse bone marrow by flow cytometry, including lymphoid-primed multipotent progenitors (LMPPs), all-lymphoid progenitors (ALPs) and B cell-biased lymphoid progenitors (BLPs). Single cell RNA-seq was performed as described previously (Picelli et al. (2014) Nature protocols 9:171).为明确Mbd3/NuRD在淋巴细胞生成（lymphopoiesis）中的作用，本研究采用单细胞RNA测序（single cell RNA-seq）技术，分析了单个Mbd3敲除型与对照型淋巴祖细胞的基因表达情况。实验整体设计：通过流式细胞术从小鼠骨髓中分离得到单个Mbd3缺陷型及对照淋巴祖细胞，涵盖淋巴定向多能祖细胞（lymphoid-primed multipotent progenitors, LMPPs）、全淋巴祖细胞（all-lymphoid progenitors, ALPs）以及B细胞偏向性淋巴祖细胞（B cell-biased lymphoid progenitors, BLPs）。单细胞RNA测序的实验操作参照既往发表的方法（Picelli等，2014，Nature Protocols，9卷：171页）。