Population RNA sequencing of Ly6D+ and TN EPLM subpopulations as well as Pro-B cells

Nowadays, there is an active debate regarding the heterogeneity of apparently phenotypically homogenous progenitors having different multiple lineage potentials. It has been previously characterized a B220+ CD117int CD19- NK1.1- uncommitted and multipotent haematopoietic progenitor with combined lymphoid and myeloid developmental potential, the so called Early Progenitor with Lymphoid and Myeloid potential (EPLM). This project is devoted to unravelling whether the EPLM is truly multipotent or whether it is composed by a mixture of cells with distinct lineage biases. Based on the expression of the cell-surface markers Ly6D, SiglecH and CD11c, the EPLM can be subdivided into at least four major subpopulations. By interrogating the transcriptome of the subpopulations (bulk RNA sequencing), we aim to unravel their genetic signatures and to identify the key genes driving their precursor-product relationship. Overall design: Total RNA was extracted from ex-vivo sorted hematopoietic progenitors (from Flt3Ltg mice) using TRIzol-based method. Transcriptome profiles were generated by sequencing, in quadruplicates, using NextSeq 500.