Hematopoietic differentiation in healthy donors and GATA2 deficiency. (A) A tSNE plot of single-cell gene expression of healthy donors. (B) Reconstruction of the hematopoietic hierarchy pseudo-time ordering in Monocle. Cells are colored by types (HSC, MEP, GMP, ETP, and ProB). (C) Average expression of characteristic genes for lineages are highlighted in a differentiation tree. HSC genes: SPINK2, CRHBP, MEIS1, and MLLT3; MEP genes: GATA1, HBD, TFRC, UROD, NFIA, and KLF1; myeloid progenitor genes: MPO, CEBPA, ELANE, IRF8, and LGALS1; lymphoid progenitor genes: DNTT, CXCR4, CD79A, BLNK, IGLL1, and EBF1. The characteristic gene list is from our previous findings using scRNA-seq and validated by qualitative polymerase chain reaction and literature mining (supplemental Figure 2). (D) Expression of GATA2 and GATA1 highlighted in the differentiation tree. (E) Dynamic expression of GATA2 and GATA1 along pseudo-time of HSC differentiation into MEP. The x-axis shows pseudo-time ordering from HSC to MEP estimated by Monocle 2. The y-axis shows normalized gene expression by scaleData function in Seurat package. (F) Top 20 coexpressed (pink) and anticoexpressed (blue) genes with GATA2 and GATA1 (yellow). Red and blue lines connecting the genes indicate the top positive and negative correlations, respectively. Same plots showing positive and negative correlations of all values are in supplemental Figure 4.