Functional association between cell proliferation and lineage restriction. (A) Three-dimensional FLE plot of 10 385 cells from the A cluster in Figure 6D. Cells are color coded by their respective cell cycle phases, with transitions between phases indicated by dashed red loops. (B-C) Three-dimensional FLE plot, with cells color coded based on their surface phenotypes (B) and gene expression levels (C) (the scale represents log2-transformed values). (D-F) Three-dimensional FLE plot (from a distinct angle), with cells color coded by their cell cycle phases (D), phenotypes (E), and gene expression (F), respectively. (G) Experimental design for assessing cell cycle progression and differentiation outputs of CD34⁺RA^–C^– progenitors treated with 100 nM wortmannin (Sigma-Aldrich, catalog no. 681675). The inhibitor is replenished every 2 days to account for its short half-life. (H) Number of output cells per 100 input cells analyzed on day 6. Each bar represents the mean ± SEM, calculated from 6 experimental replicates derived from 2 independent CB pools. (I-J) Numbers of CD34⁺RA^–C^– (I) and CD34^– (J) output cells detected across successive CFSE fluorescence peaks, reflecting the number of completed cell divisions. Each point represents the mean ± SEM from the same experiments as shown in panel H. ∗P < .05, determined by t tests. (K) A proposed model of the process of cell fate restriction into lymphoid and myeloid lineages from bipotent human progenitors. Ctr, control; DMSO, dimethyl sulfoxide; FLE, force-directed layout.