Differential expression of signaling pathway modulators in Gata2-dependent and -independent HPCs. (A) Flow cytometric sorting gates for isolation of E10.5 AGM G2V CD31⁺cKit^intVenus⁻ (gray) and CD31⁺cKit^intVenus⁻ (green) HPCs used for RNA sequence analysis. Gene Expression Omnibus data accession number is GSE76254. (B) Gene enrichment analysis for Ras signaling pathway genes. Bar graphs of fragments per kilobase million (FPKM) values obtained from RNA sequence analysis of CD31⁺cKit^intVenus⁻ (gray bar) and CD31⁺cKit^intVenus⁺ (green bar) AGM cells for (C) Ras pathway and cyclic AMP response element-binding protein (CREB) and CREB-binding protein (CBP) transcription factor genes and (D) Notch pathway genes. (E) Bar graphs of FPKM values obtained from RNA sequence analysis for a selection of chemokine receptor/ligand genes (see Kierdorf et al³⁷ ; these genes were down-/upregulated in YS EMPs compared with adult microglia [AM]). (F) Representative FACS plots demonstrating frequency of EMPs in the Venus⁺ fraction, as defined as Sca1⁻cKit⁺CD41⁺CD16/32⁺, in YS and AGM of E10 (left) and E11 (right) G2V embryos. Numbers indicate the percentages of gated cells within the parental cell population.