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. 2023 Nov 1;221(1):e20231035. doi: 10.1084/jem.20231035

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© 2023 Solomon et al.

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Figure 4. — Label retention properties reveal a hierarchy within the MPP compartments. (A) Schematic describing the Tet-ON inducible H2B-GFP mouse model used to observe long-term divisional kinetics and history of HSC, MPPs, and lineage-committed progenitors. (B) Percentage ± SD of GFP maximum fluorescence intensity in BM HSPC populations of H2B-GFP reporter mice after 6–10-wk chase. (C) Representative FACS plots showing GFP fluorescence in BM HSPCs of H2B-GFP reporter mice after 6-, 8-, 10-, or 20-wk chase. Insert shows equal H2B-GFP labeling in LSK Flt3⁻, MPP4, and myeloid progenitor (MP) compartments after 2-wk pulse. (D) Representative FACS plots showing phenotypic heterogeneity within MPP4 based on CD48 and CD62L. The graph indicates the relative percentage ± SD of each subset in the MPP4 compartment. (n = 9 from three independent experiments). (E) Representative FACS plots showing Ly6A/E (Sca1) and H2B-GFP expression in MPP4 subset defined by CD48 and CD62L. Representative of three independent experiments. (F) Competitive BM transplantation assays in conditioned recipient using CD48⁻CD62L⁻, CD48⁺CD62L⁻ ⁺, and CD48⁺CD62L⁺ MPP4 subsets. Graphs indicate percentage ± SD of total donor chimerism in PB and percentage ± SD of myeloid cells in donor-derived cells over time. n = 3/5/3 per group from one experiment. One-way ANOVA (B) or two-way ANOVA (F) with Tukey’s post hoc test: *, P ≤ 0.05; **, P ≤ 0.005; ****, P ≤ 0.0001.