Fig. 5

Distinct erythroid-associated and megakaryocyte-associated transcriptional lineage-priming in MEP subpopulations. a Population 1 (green) contained cells with residual CSF3R, FLT3/CD135, and SOCS3 expression and lowest GATA1 and GATA2 expression, suggesting that this population comprises progenitors earlier in the hematopoietic hierarchy than populations 2 and 3 and more closely related to CMP. Expression of myeloperoxidase (MPO) was only detected in five of 681 cells, indicating minimal contamination of the FACS-isolated MEP cells with CMP or other myeloid lineage cells, in which MPO is strongly positive [20]. b The highest levels of expression of erythroid genes, including KLF1, TMOD1, ANK1, LEF1, and ADD2 were observed in Population 2 (purple). c The highest levels of expression of megakaryocyte genes, including VWF, FLI1, NFIB, TGFB1, and LOX occurred in Population 3 (orange). Each chart shows a bee-swarm plot where each dot represents the gene expression of an individual cell, with a box plot overlaid. Significance values are shown for q-values for KS test with FDR correction between populations: *-q <0.05; **-q <0.01; ***-q <0.001; NS-q >0.05. d Heatmap showing correlation of expression of selected erythroid and megakaryocytic genes within single cells. Color-coding: Orange box, megakaryocyte gene set; purple, erythroid; yellow, both megakaryocyte and erythroid; green, genes associated with pre-MEP phenotype. e, f Representation of Spearman correlation coefficient between selected genes in populations 2 (Fig. 5e) and 3 (Fig. 5f), respectively. Blue edges denote positive correlation and red edges denote negative correlation. Edge thickness is a function of correlation magnitude