Figure 6.

Ebf1 controls lymphoid/myeloid balance in HSPCs, and its loss causes increased myeloid cells in the BM. (A and C) Annotated UMAP projection of scRNA-seq landscape derived from Ebf1^WT and Ebf1^KO LK and LSK populations. Biological replicates n = 4. (B and D) Log-normalized Ebf1 expression for Ebf1^WT cells shown in A and C, respectively. (C) Cells from Nestorowa data (Nestorowa et al., 2016; colored) were embedded into the Ebf1^WT/Ebf1^KO landscape (gray) with annotated immunophenotypic populations. (E) Log-normalized expression of Ig and lymphoid-associated DE genes in Ebf1^WT and Ebf1^KO cells of selected clusters. Each dot represents the mean expression per mouse (four mice in total). (F) Number of DE genes between Ebf1^WT and Ebf1^KO cells per cluster. (G) DoT scores calculated using DE genes in cluster 4 between Ebf1^WT and Ebf1^KO cells in the context of the LK/LSK landscape. Mean expression in cluster 4 was used as the point of origin. Red indicates a shift toward that cell fate and blue indicates a shift away from that cell fate. (H) Violin plots showing the gene expression score of G2/M cell cycle–associated genes in Ebf1^WT and Ebf1^KO cells in selected clusters. (I) UMAP visualization of the expression of Myc and selected Myc target genes in Ebf1^WT and Ebf1^KO cells. (J) Boxplots showing the frequency of myeloid cells (CD11b⁺) in the BM. Ebf1^WT n = 18, Ebf1^KO n = 16, Ighm^WT and Ighm^Tm1 n = 11. Statistical significance was determined by Mann–Whitney U test. Data are from >2 independent experiments. LK defined as Lin-cKit⁺ cells. ESLAM, CD45⁺EPCR⁺CD48⁻CD150⁺ HSCs; Meg, megakaryocyte; Ery, erythrocyte; Neu, neutrophil; Mono/DC, monocyte/dendritic cell; Ly, lymphocyte.