The Ets transcription factor Er71 is an important regulator of endothelial and hematopoietic development during mammalian embryogenesis. However, the role of Er71 in adult hematopoiesis has remained unknown. We now first show that conditional deletion of Er71 in the hematopoietic system of adult mice results in a marked reduction (55%) in the number of hematopoietic stem cells (HSCs) that is likely due to increased cell death. Bone marrow transplantation (BMT) experiments further confirmed that Er71 is required for repopulation of HSCs. In addition, Er71^+/− mice exhibited a slight decrease (37%) in the number of HSCs than those of Er71^+/+ mice, indicating that the function of Er71 in HSC maintenance is dependent on gene dosage. Moreover, Er71 was shown to be required for Tie2 expression, which contributes to HSC maintenance. Our results thus suggest the role of a single transcription factor in controlling HSCs through regulation of Tie2 expression in adult animals.