Hmga2 is a RUNX1 target gene in HSPCs. (A) Venn diagram showing the overlap between the common differentially expressed genes in Runx1^Δ/Δ and RUNX1SF HSPCs (n = 59)¹⁰  and the genes associated with RUNX1 ChIP-seq peaks. (B) Relative expression of Hmga2 in wild-type, Runx1^Δ/Δ, and RUNX1SF HSPCs based on microarray data published in Matsuura et al.¹⁰  (C) Validation of upregulation of Hmga2 in microarray results by RT-qPCR using cDNA from wild-type, Runx1^Δ/Δ, and RUNX1SF HSPCs. RNA extraction and RT-qPCR were performed from samples of at least 3 independent batches of mice. HMGA2 protein levels by intracellular flow cytometry are shown in (D) LSK, (E) GMP, and (F) CMP cell populations between wild-type and Runx1^Δ/Δ mice (n = 5 each). (G) Location of RUNX1 ChIP-seq peaks relative to the Hmga2 gene locus on chromosome 10. IgG control and one RUNX1 ChIP-seq replicate are shown. (H) Confirmation of RUNX1 occupancy regions in relation to the Hmga2 gene locus by ChIP-qPCR. RUNX1 ChIP was compared relative to IgG ChIP and normalized to a negative control region. Data represent 3 replicates of ChIP followed by qPCR.