Abrogation of LNK enhances the engraftment of transplantable HSCs in the bone marrow. CB-derived CD34⁺ HSPCs were transduced with lentiviral shRNAs to Luc or LNK and transplanted into NBSGW mice as described in Figure 1. (A) Mean percent mCherry⁺ in the engrafted CD34⁺ compartment in the BM. (B) Mean percent mCherry⁺ in the engrafted CD34⁺CD38⁻ compartments in the BM HSCs (CD34⁺CD38⁻CD45RA⁻CD90⁺), multipotent progenitors (MPPs) (CD34⁺CD38⁻CD45RA⁻CD90⁻), and multilymphoid progenitors (MLPs) (CD34⁺CD38⁻CD45RA⁺CD90^lo/−). (C) Representative HSPC flow cytometric analysis of the transplanted BM. (D) Proportion of HSPC subpopulations in the mCherry⁺CD34⁺CD38⁻ compartment of engrafted BM. N = 23 shLuc, n = 9 shLNK #1, n = 7 shLNK #2, and n = 8 shLNK #3 transplanted mice. The data are pooled from biological replicates n = 8 CBs for shLuc, 3 CBs for shLNK #1, 2 CBs for shLNK #2, and 3 CBs for shLNK #3. (E) After 16 to 27 weeks, total BMs from shLuc and shLNK #3 primary recipients were transplanted into secondary NBSGW recipients. Mean mCherry⁺ percent in hCD45⁺ PB at week 12 is shown. N = 9 shLuc, n = 8 shLNK #3 mice. In all relevant panels, each symbol represents an individual mouse; bars indicate mean values; error bars indicate plus or minus SEM. *P < .05; **P < .01; ***P < .001; as determined by 2-way ANOVA followed by Dunnett’s multiple comparisons to shLuc. Secondary xenotransplantation was analyzed by 2-tailed Student t test.