Mobilized HSPC transplantation defects are divisional history independent. (A) Schematic of BM/PB chimeric HSPC transplantations. LSKCD48⁻ cells were sorted from the bone marrow of 34/H2B mice (CD45.2⁺) chased with dox for 12 weeks, based on label dilution. These cells were mixed together to mimic the label dilution ratios of LSKCD48⁻ cells found in mobilized PB 3 hours after G-CSF treatment (Figure 2C). This BM HSPC mix was then further mixed at equal ratios with LSKCD48⁻ cells sorted from mobilized PB of UBC-GFP mice (CD45.2⁺). This BM/PB HSPC mix was then competitively transplanted into CD45.1⁺ recipient mice. (B) Plot displaying GFP intensities of PB derived from UBC-GFP and 34/H2B mice. (C-D) PB- and BM-derived HSPC contribution to PB in primary (C) and secondary (D) transplantations. (E) Gating strategy for the BM HSPC compartment. (F-G) PB and BM derived contributions to the regeneration the BM HSPC compartments. In secondary transplant, regeneration in the HPC-2 and HSC compartments was exclusively confined to BM-derived cells, which prevented calculation of statistical significance between the BM- and PB-derived groups; not determined (n.d.). Data are represented as mean ± SEM of 8 and 9 mice for primary and secondary transplants, respectively. *P < .05, **P < .01, ***P < .001 by paired t test.