Fig. 4

DAB2 regulates HSPC function in vivo. a Colony-forming unit (CFU) analysis of constitutively expressing DAB2 (DAB2-CE) marrow (mean ± SEM, n = 7). b Primary CFU marrow was replated in equal proportions per condition, normalized to the number of input cells, to generate secondary CFUs (mean ± SEM, n = 7). c Schematic of the transplant experiments shown in d through h. CBC complete blood counts, LDA limiting dilution assay. d Recipient mice were injected with equal numbers of Vector-YFP and DAB2-GFP marrow in a competitive assay (mean ± SEM, week 4 n = 5, week 8–12 n = 9, week 20 n = 8). Peripheral blood chimerism was assessed every 4 weeks for 20 weeks. e Peripheral blood analysis at 20 weeks (mean, n = 8) to assess the percentage of donor granulocyte–monocyte cells (GM; CD45.1⁺Mac1⁺ and/or CD45.1⁺Gr1⁺), B cells (CD45.1⁺CD19⁺) and T cells (CD45.1⁺CD3⁺). f Bone marrow was harvested at 20 weeks to assess GFP/YFP chimerism (mean, n = 8) of LSK (Lin⁻Sca1⁺c-Kit⁺) and myeloid progenitors (CMP, Lin⁻Sca1⁻c-Kit⁺CD34⁺CD16/32^lo; GMP, Lin⁻Sca1⁻c-Kit⁺CD34⁺CD16/32^hi; MEP, Lin⁻Sca1⁻c-Kit⁺CD34⁻CD16/32^lo). g Results from f were reanalyzed by gating on the transduced population to determine the proportion of specific progenitors within each of the GFP or YFP compartments. h Secondary limiting dilution assays were performed using marrow from long-term engrafted mice in d (20 weeks post-competitive DAB2-GFP/Vector-YFP transplant) and engraftment was evaluated at 16 weeks post-secondary transplant. Shown is a log-fraction plot of the limiting dilution model. The slope of the line is the log-active cell fraction. The dotted lines give the 95% CI