Supplementary Figure 6: Notch signalling does not mediate competition between endothelial cells during intersegmental blood vessel sprouting.
From: Endothelial Notch signalling limits angiogenesis via control of artery formation
(a) Transplantation scheme. Tg(kdrl:EGFP)s843 marks donor cells that were either injected with a ctr or dll4 MO in green. Tg(kdrl:Hsa.HRAS-mcherry)s916 marks host vasculature in red. (b) Representative image of mosaic embryos at 32 hpf. (c) Quantification of contribution of ctr MO (n = 19 mosaic embryos) or dll4 MO (n = 23 mosaic embryos) injected ECs to different positions within the trunk vasculature (d) Transplantation of dll4 mutant cells reveals no preference for cell position within trunk blood vessels. Number of embryos with indicated distribution of transplanted cells indicated (n = 25 dll4+/− to WT mosaic embryos; n = 10 dll4−/− to WT mosaic embryos in 3 independent experiments). (e) Transplantation scheme. Tg(fli1a:EGFP)y1 marks donor cells that were either wt, hetero- or homozygous for mib in green. Tg(kdrl:Hsa.HRAS-mcherry)s916 marks host vasculature in red. (f) Representative image of mosaic embryos at 32 hpf. Arrowheads mark donor cells at tip positions. (g) Quantification of contribution of mib sibling (n = 17 mosaic embryos in 3 independent experiments) or homozygous mib mutant (n = 12 mosaic embryos in 3 independent experiments) ECs to different positions within the trunk vasculature. Images are representative of three independent experiments. (h) Still images of video 1 at indicated time points of ISV sprouting. Tg(fli1a:nEGFP)y7 marks EC nuclei in green. Numbers delineate individual ECs or their progeny. Out of 10 ISVs observed, we only detected one shuffling event (white arrowhead in g). (i) Still images of video 2 at indicated time points of ISV sprouting. Tg(fli1a:nEGFP)y7 marks EC nuclei in green. Numbers delineate individual ECs or their progeny. Out of 10 ISVs observed, we did not detect EC shuffling. (n = 2 embryos, 2 independent experiments). Scale bars, 50 μm. aISV-arterial intersegmental vessel; ctr-control; DA-dorsal aorta; DLAV-dorsal longitudinal ananstomotic vessel; hpf-hours post fertilization; MO-morpholino; PCV-posterior cardinal vein.
