LNK depletion enhances engraftment of umbilical CB HSCs in PB of xenotransplanted mice. (A) Experimental scheme of lentiviral transduction and xenotransplantation assay. Primary human CD34⁺ HSPCs were isolated from CB and transduced with lentivirus expressing miR30-based short hairpin RNAs (shRNAs) to either Luc or LNK and coexpress mCherry as a fluorescent marker. Subsequently, transduced HSPCs were transplanted into immunodeficient NBSGW mice. (B) mCherry⁺ percentages in the hCD45⁺ PB of transplanted mice were examined every 4 weeks, up to 16 weeks, and percent mCherry⁺ within hCD45⁺ of individual mice in the PB over time is shown, represented by each solid line. Gray bars indicate the range of shLuc and shLNK mCherry⁺ transduction rates at time of transplant. Each graph tracks the reconstitution of an individual CB. (C) Mean percent mCherry⁺ within hCD45⁺ PB cells at different time points posttransplant is shown. 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 (shLuc vs shLNK #1 and #2) or 2-tailed Student t test (shLuc vs shLNK #3). 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. Ns, not significant