Single-cell analysis reveals that gata2b^−/− KM cells are overrepresented in lymphoid lineage clusters and reduced in erythroid and myeloid lineage clusters compared with WT. (A) Split uniform manifold approximation projection (UMAP) of WT and gata2b^−/− cells with cluster indication of enriched (arrow up) or reduced (arrow down) cell clusters in gata2b^−/− cells. (B-E) Pooled WT and gata2b^−/− UMAP feature analysis with gradual gene expression in shades of blue. Expression pattern of granulin1 (grn1) (B), cluster of differentiation 37 (CD37) (C), hemoglobin β adult (hbba1) (D), and gata2b (E). (F-I) Violin plots representing the expression levels of genes within the different clusters; each dot represents expression in 1 cell. Fli-1 proto-oncogene (fli1a) (F), meis homeobox 1b (meis1b) (G), gata2b (H), and GFP (I), indicating CD41:GFP^low cells. (J) Volcano plot comparing HSPC1s vs HSPC2s. At the left of the y-axis are genes in HSPC1s with an average logarithmic fold change less than −0.25, and to the right are genes with a logarithmic fold change >0.25 compared with HSPC2s. (K) Lineage differentiation trajectory depicted on UMAP with WT cells in blue and gata2b^−/− cells in pink. (L) Pseudotime analysis assuming HSPC1s as a starting point. (M) Quantitation of proportions of distribution between WT and gata2b^−/− cells in the different clusters. Significant differences are indicated in red. (N-Q) Pseudotime analysis of gene expression in lineage trajectory analysis of gata2b (N), CD37 (O), grn1 (P), and hemoglobin β adult (hbba1) (Q). FDR, false discovery rate; n.s., not significant.