Figure 5
Figure 5. RUNX1 directly controls transcription of integrin β3 (CD61) in the HE. (A) (Left) RPKM mapped reads expression values of CD61, CD41, and CD51 genes in Runx1wt and Runx1ko HE. (Right) Fold change of RPKM values of CD61, CD41, and CD51 genes in Runx1wt HE normalized against Runx1ko. *P < .05. (B) qPCR for CD61 mRNA expression levels in non-HE (cKIT−, TIE2−, CD41−,CD45−, and TER119−) and HE (cKIT+, TIE2+, CD41−, CD45−, and TER119−) enriched populations sorted from E7.5 and E8.5 heterozygous (HET) (RUNX1RFPp/+) and KO (RUNX1RFPp/RFPp) embryos. Data are normalized against the HET HE population. (C) Luciferase assay with the full length and truncated +16 kb CD61 cis-regulatory element cloned upstream of the CD61 promoter. The “2R” depicts the 2 active RUNX1 binding motifs. The activity of all luciferase vectors was tested in iRunx1Runx1−/− HE 24 hours postdox induction. Values were normalized against “no dox” treatment. *P < .05. (D) cKIT/TIE2 and CD61/CD41 FACS plots of iRunx1Runx1−/− HE cells in the presence or absence of dox. HP, hematopoietic progenitors.

RUNX1 directly controls transcription of integrin β3 (CD61) in the HE. (A) (Left) RPKM mapped reads expression values of CD61, CD41, and CD51 genes in Runx1wt and Runx1ko HE. (Right) Fold change of RPKM values of CD61, CD41, and CD51 genes in Runx1wt HE normalized against Runx1ko. *P < .05. (B) qPCR for CD61 mRNA expression levels in non-HE (cKIT, TIE2, CD41,CD45, and TER119) and HE (cKIT+, TIE2+, CD41, CD45, and TER119) enriched populations sorted from E7.5 and E8.5 heterozygous (HET) (RUNX1RFPp/+) and KO (RUNX1RFPp/RFPp) embryos. Data are normalized against the HET HE population. (C) Luciferase assay with the full length and truncated +16 kb CD61 cis-regulatory element cloned upstream of the CD61 promoter. The “2R” depicts the 2 active RUNX1 binding motifs. The activity of all luciferase vectors was tested in iRunx1Runx1−/− HE 24 hours postdox induction. Values were normalized against “no dox” treatment. *P < .05. (D) cKIT/TIE2 and CD61/CD41 FACS plots of iRunx1Runx1−/− HE cells in the presence or absence of dox. HP, hematopoietic progenitors.

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