Preservation of in vitro determined early B and NM precursor phenotypes in CB-derived xenografts. (A) FACS gating used to define RA^–C^–, RA⁺C^– and C⁺ phenotypes in unmanipulated CD34⁺38med71^–10^– CB cells. (B) Clonal output frequencies of the 3 input phenotypes isolated from unmanipulated CD34⁺ CB cells. Each bar shows the mean ± SEM of values pooled from 4 different experiments with 150 to 340 single cells tested per input subset. The statistical significance of differences in clone distribution between input phenotypes was assessed using t tests with Holm correction (∗P < .05). (C) Experimental design used to analyze clonal outputs of RA^–C^–, RA⁺C^–, and C⁺ phenotypes obtained from the bone marrow (BM) of NRG-W41 mice transplanted with CD34⁺ CB cells (4 experiments). (D) FACS gating used to detect the phenotypes generated in panel C. (E) Clonal output frequencies of 3 input phenotypes isolated from the BM of engrafted NRG-W41 mice. Each bar shows the mean ± SEM of values pooled from all experiments with 260 to 550 total single cells tested per input subset. The statistical significance of differences in clone distribution between input phenotypes was assessed using t tests with Holm correction (∗P < .05). (F) Size of clones generated from single input cells isolated from the different sources described above. Clones (defined as ≥5 human cells) pooled from all experiments shown above were grouped by the input phenotype (left) or lineage output type (right). “n.d.” represents signals below the limit of detection (5 cells). (G) Experimental design for the OP9-DLL4 coculture assay of clonal pro-T– and NM-lineage outputs in sorted single cells from each of the 3 input phenotypes. (H) Frequency of clonal output types detected in the OP9-DLL4 clonal assays. Values shown are mean ± SEM pooled from 3 experiments, with 180 to 240 cells tested per input phenotype. The statistical significance of differences in clone distribution between input phenotypes was assessed using Wilcoxon tests with Holm correction (∗P < .05).