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Blood, Vol. 113, Issue 22, 5680-5688, May 28, 2009
DLL1-mediated Notch activation regulates endothelial identity in mouse fetal arteries
Blood Sörensen et al.
113: 5680
Supplemental materials for: Sorensen et al
Files in this Data Supplement:
- Figure S1. Peripheral vessels show the same arterial phenotype as vessels at the axial position of the outflow tract (JPG, 814 KB)
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Mutant vessels from the head and the abdominal region showed loss of Notch activation (A, B; I, J) as well as downregulation of arterial markers (C–H, K–P).
- Figure S2. Notch signaling components in the vascular endothelium (JPG, 317 KB)
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(A) Expression of Notch1-4 (a–h), Dll4 (i) and Jagged1 (j) was detected in Dll1 mutant embryos at E15.5, whereas expression of the Notch target Hey2 appeared significantly reduced (m, n). Maniac Fringe appeared upregulated in the endothelium of bigger vessels (p, p′) in Dll1 hypomorphs. a′ to p′ show magnifications of the boxed regions shown on a–p. (B) Notch transactivation assay: Notch signaling is highly activated, when CHO cells stably expressing DLL1 or DLL4 were cocultivated with HeLa-N1 cells. Transient coexpression of DLL1 and DLL4 in CHO cells or coexpression of MFNG with one of the ligands did not affect Notch activation. Transient expression of MFNG in HeLa-N1 cells strongly increased DLL1 and DLL4 mediated Notch activation. Labels on the right of the legend indicate transiently expressed components. Bars indicate standard deviations. Expression of components that were transiently transfected was verified by Western blot analyses of cell lysates and showed comparable expression levels.
- Figure S3 (JPG, 439 KB)
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(A) Dll1 hypomorphic and endothelial specific mutants show a significantly reduced vessel lumen at E18.5. At E15.5 the difference did not reach statistical significance. The vessel lumen was measured on HE stained paraffin sections using Image J. Data represent mean values of 6 embryos per embryonic stage and genotype. Bars indicate standard deviation. (B) Ink injections in the carotid artery of E17.5 wildtype and mutant embryos. In some mutants the vessel network appeared unorganized in the head region of Dll1 endothelial-specific mutants, but no consistent defects could be found. (C) Retina preparations of P6 and P15 wildtype (a, b) and endothelial-specific Dll1 knock out mutant pups (f, g) stained with IsolectinB4 (blue), anti-Desmin (green) and -SMA (red) revealed no significant difference in angiogenic network formation. The postnatal induced DLL1 deletion was confirmed by anti-DLL1 staining (c, h, green). PECAM staining of P15 brains of wild type (d, e) and endothelial-specific Dll1 mutant pups (i, j) also did not show obvious abnormalities in mutants.
- Figure S4. Dll1 (a–f) and Dll4 (g–l) expression in embryonic vessels detected by in situ hybridization (JPG, 292 KB)
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Prior to E13.5 only Dll4 transcripts were detected in arterial vessels.
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