Transcriptional deregulation in EZH2-deficient lymphocyte progenitors

This goal of this study was to identify genes that are deregulated in the absence of EZH2 in early lymphocyte progenitors. We examined gene expression by RNA-sequencing in sorted CLPs (Lin-CD117int CD127+ CD135+), pro-B cells (B220+CD19+CD43+), DN3 cells (Lin- CD25+ CD117-), splenic NK cells (Lin-NK1.1+DX5+) and bone marrow ILC2 cells (Lin- Sca1+CD127+) from Ezh2fl/fl Il7racre/+ and Il7racre/+ control mice. Reads were aligned to the mm10 reference genome by Tophat2.1.0. Reads were assigned to genes using the htseq-count tool from HTSeq v 0.6.1 and gene annotations from Ensembl release 78. Differential expression was calculated across 2-3 independent replicates by EdgeR. We found that CLPs, ILC2s, and splenic NK cells maintained their normal transcriptional programs despite loss of EZH2. In contrast, loss of EZH2 caused over 1000 genes to be deregulated in pro-B and DN3 cells indicating that EZH2 is required for transcriptomic stability in adaptive, but not innate lymphocyte progenitors. Expression profiling analysis of CLPs, pro-B, DN3, splenic NK, and ILC2 in Ezh2fl/fl Il7racre/+ mice and Il7racre/+ controls generated by RNA-sequencing. 2-3 independent replicates per sample.