Global mRNA profiling (RNA-seq) across mouse hematopoietic hierarchy

Epigenetic regulation exerts a crucial role in maintaining the balance between self-renewal and differentiation of hematopoietic stem cells (HSCs), thereby sustaining normal hematopoiesis. Although emerging evidence has demonstrated the contribution of RNA N4-acetylcytidine (ac4C) modification and its catalytic enzyme Nat10 to numerous biological processes, particularly tumorigenesis, the landscape and function of ac4C in normal hematopoiesis remain unexplored. In this study, we characterize the landscape of ac4C across the hierarchy of hematopoietic stem and progenitor cells (HSPCs) using RNA-seq and reveal a key role of Nat10 in the maintenance of HSCs. Overall design: To understand the role of epigenetic regulation in the establishment and maintenance of HSC fate decisions, we sorted distinct hematopoietic populations including long-term HSCs (LT-HSCs), short-term HSCs (ST-HSCs), multipotent progenitors (MPP), common myeloid progenitors (CMP), granulocyte-macrophage progenitor (GMP), megakaryocyte/erythroid progenitors (MEP), and the mature cells from the bone marrow (BM) using fluorescence-activated cell sorting (FACS), followed by RNA sequencing (RNA-seq) analysis.