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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 3. — Slow- and fast-cycling MPP4 display specific functional properties. (A) Phospho-flow analysis measuring ERK phosphorylation after 60 min stimulation with Flt3L (50 ng/ml). The left panel shows mean fluorescence intensity (MFI) for pERK displayed as a percentage ± SD relative to Flt3-treated LSK Flt3⁻ early hematopoietic populations (set at 100). The right panel shows a representative FACS plot showing pERK detection in CD62L^+/− MPP4 cells. n = 3, data from two independent experiments. (B) Single-cell tracking measuring the kinetics of the first division for cultured CD62L^+/− MPP4 cells compared to HSCs. The left panel shows cloning efficiency. The right panel shows the cell division kinetics for each subset (60 cells/plate, n = 2). (C and D) Competitive BM transplantation assays in conditioned recipients using non-limiting (C) and limiting (D) number of CD62L^+/− MPP4 cells. Graphs indicate percentage ± SD of total donor chimerism in PB and percentage ± SD of myeloid and B/T lymphoid cells in donor-derived cells over time. (C) n = 6/group from one experiment. (D) n = 6–4/group, representative of two independent experiments. One-way ANOVA with Tukey’s post hoc test (A) or two-way ANOVA with Sidak’s post hoc test (C and D): *, P ≤0.05; **, P ≤ 0.005; ****, P ≤ 0.0001. WBM: whole BM; IV: intravenous.