Supplementary Figure 5: Gene expression in LSKs and GMPs from WT and ACKR1-deficient mice.

(a) Microarray heatmap of genes expressed in LSKs and GMPs from wild-type (WT) and ACKR1-deficient (KO) BMs. Each row represents a gene and columns show individual cell populations, in duplicates. Two clusters defined by hierarchical clustering analysis reflect specific gene expression in LSK and GMP. The levels of expression of specific LSK and GMP enriched genes were similar in WT and KO BM. (b) Left, blue: Comparative gene expression in neutrophils (PMNs) and GMPs from Immgen dataset (www.immgen.org). The expression of the 444 genes was higher in PMNs vs. GMPs on average >10 times. Three genes with the highest increase, Ltf, Ly6G and Mmp8, were expressed ca. 100 times more in PMN, than in GMP. Right, red: comparative gene expression in GMPs from WT vs. KO BM expressed as fold change. The level of expression of the overwhelming majority of genes was the same in GMPs from WT and KO BMs (mean fold change 1) but some were expressed orders of magnitude higher in KO GMPs. The transcripts for three most overexpressed genes Rentlg, Camp and Ngp, were in excess of 1000 times higher in KO vs. WT GMPs. These genes were not highly upregulated during the differentiation of GMPs towards PMNs (left panel, blue). This excluded a potential contamination of the GMP with PMNs and was consistent with the induction of a specific transcriptional program of a subset of neutrophil specific genes (also see Fig. 3c) in GMPs rather than only their conventional differentiation to PMNs. (c) Microarray heatmap from Fig. 3a with the list of genes. (d) Microarray heatmap from Fig. 3b with the list of genes.