Figure 3. Production of dendritic cells (DCs) and megakaryocyte progenitors (MkP) by hematopoietic stem and progenitor cells (HSPCs) after erythropoietin (EPO) exposure and transplantation.

In addition to the analysis of barcodes in the erythroid (E), the myeloid (M), and the B-cell (B) lineage, the DC lineage in spleen and MkP in bone marrow were added. (a) Percentage of barcoded HSPCs producing DC in the different HSPC categories (classification as in Figure 2 based on the M, E, and B lineage only using a 10% threshold; the DC-only category was added). (b) The percentage of the DC lineage produced by the barcodes categorized by bias as in (a). (c, d) Representations as in (a, b) for barcode detection in MkP. Data is derived from a cohort with detailed myeloid sorting. The myeloid lineage was merged according to the percentage of total donor myeloid each subset contributed as in Figure 2—figure supplement 1a to allow classification as in (a, b) based on the M, E, and B lineage only using a 10% threshold. The MkP-only category was added. Shown are values from several animals (a, b, n = 5 for the control group and n = 2 for the EPO group; c, d, n = 3 for the control group and n = 1 for the EPO group [collected over two experiments]). For all bar graphs, mean and SD between mice are depicted. Statistical significance tested using Mann–Whitney U-test p=0.05 for (a, c). Statistical significance tested by permutation test for different subsets in (b) (see Table 1).

Figure 3—figure supplement 1. Production of macrophages (Ma), monocytes (Mo), neutrophils (Neu), eosinophils (Eo), and megakaryocyte progenitors (MkP) by hematopoietic stem and progenitor cells (HSPCs) after erythropoietin (EPO) exposure and transplantation.

Same experimental protocol as in Figure 1 but the myeloid cells were subdivided into Mo, Eo, Ma, and Neu, and MkP were sorted. (a) Gating for detailed myeloid subsets on myeloid cells. Mo were sorted as CD115⁺ cells, Eo as CD115^- SiglecF⁺ Ly6G⁺, Ma as CD115^- SiglecF^- Ly6G^-, and Neu as CD115^- SiglecF^- Ly6G⁺ cells. (b) The contribution of different cell types to the overall donor myeloid subset in the control and EPO groups. (c) Heatmaps showing the output of individual barcodes (rows) in different samples (columns) as indicated. Data is normalized by cell subset, log transformed, and clustered by complete linkage using Euclidean distance. No output is represented in black. Shown are values from several animals (b, n = 5 for the control group and n = 3 for the EPO group, c, n = 3 for the control group and n = 2 for the EPO group [collected over two experiments]). For all bar graphs, mean and SD between mice are depicted. Statistical significance tested using Mann–Whitney U-test p=0.05 for (b).