Notch progressively depletes HSC activity and numbers over time. (A) Lethally irradiated CD45.1⁺ congenic mice were reconstituted with 5-FU–treated donor Rag1^−/−CD45.2⁺ BM cells transduced with MigR1 control or activated Notch1 (ΔEGFΔLNRΔP). BM DN T-cell production (Thy-1⁺CD25⁺) was measured at 6 weeks and 13 weeks after transplantation. Cells are gated on the CD45.2⁺GFP⁺Lineage⁻ cells. (B) Scatter plot analysis of DN T-cell production by MigR1 controls and Notch mice at 6 and 13 weeks after transplantation. (C) The LT-HSC compartment (Lineage⁻Sca-1⁺c-Kit⁺CD34⁻Flt3⁻) was measured for donor reconstitution by GFP⁺ MigR1 control and Notch-activated HSCs at 6 and 13 weeks after transplantation. (D) Scatter plot analysis of donor-derived (GFP⁺) HSCs (Lineage⁻Sca-1⁺c-Kit⁺CD34⁻Flt3⁻) at 6 and 13 weeks after transplantation. Experiments were performed 3 times.