Fig. 3.

Erythroid cells present in LSKs and myeloid progenitors selectively remain in DM embryos. A, Percentages and numbers of indicated cell subsets sorted for SORT-Seq within the E14.5 LSK compartment. B, Single-cell transcriptome-based UMAP clustering of indicated genotypes. Color-coded cells correspond to indicated clusters. C, Quantification of the frequency of cells belonging to cluster 2 in each genotype, as determined by the SEURAT algorithm. D, Heatmap of handpicked genes related to differentiation of MEPs, CLPs, GMPs as well as depiction of marker genes of cluster 2 per genotype. E, GO analysis of differential genes of cluster 2 compared to the clusters, error bars denote GO scores of clusters from all embryos combined (total n = 11). F, Residence of MPP4s in each cluster, shown as the UMAP plots of SORT-Seq. Note the increased residence of MPP4s in cluster 2 in Rev1-/- and DM embryos. G, FACS representative example of CD24^high CD93^low population in Rev1^−/− and DM FL LSKs. H, Frequency of CD24^high CD93^low population by flow cytometry in FL LSKs. I, FACS gating of LSKs, containing CD24a^highCD93^low subset in WT and Klf1^NAN E18.5 embryos. J, In each graph, the bar represents the mean ± SD. WT n = 15, Klf1^NAN = 10. Quantification of CD24 MFI of LSK cells (Left) and frequency of CD24a^highCD93^low population in WT and NAN E18.5 (Middle). The graphs on the right indicate specific cells counts per 1 × 10⁻⁶ of live cells. P values were calculated using unpaired t test with Welch correction. K, Composition of the myeloid compartment in WT and DM embryos. L and M, Relative (L) and absolute (M) numbers of indicated myeloid compartment cells. In each graph, the bar represents the mean ± SD. n for L: WT n = 7, Pcna^K164R n= 5, Rev1^−/− n = 6, Pcna^K164R Rev1^−/− n= 5. n for M: WT n = 7, Pcna^K164R n= 5, Rev1^−/− n = 9, Pcna^K164R Rev1^−/− n= 4. P values were calculated using unpaired t test with Welch’s correction.