Table S1. Notch signalling fails to rescue arterial gene expression in embryos treated with a
VegfR inhibitor
Embryos received from crosses of TG(hsp70:gal4) and TG(UAS:notch1a-intra) double
transgenic parents were treated with 5 µM VegfR inhibitor or 0.04% DMSO from 9% epiboly (9
hpf). At the 17s (17 hpf) stage, the embryos were heat-shocked to induce NICD expression.
Embryos were collected at 24 hpf and arterial gene expression was analysed. The number of
embryos with efnb2a or notch1b expression is given as a fraction of the number of embryos
analyzed.
Treatment
DMSO, 0.04%
VegfR inh., 5 µM VegfR inh., 5 µM
Gene
–
–
NICD induced
efnb2a
54/54
0/61
7/245
notchb1
13/13
0/20
2/47
Figure S1. Notch receptor, but not Notch ligand gene expression and ISV sprouting are
affected in hey2 morphants
Expression of notch5 in the DA (black arrow) is lost in hey2 morphants (a,b), while the
expression of the gene for the Notch ligand DeltaC is retained in the DA (c,c’,d,d’). DeltaC
expression appears to be missing in the position of the ISVs (red arrow in c,d, missing expression
in 44/64 embryos). Injection of the hey2 MO into fli1-egfp transgenic embryos that express
EGFP in all endothelial cells1 revealed that ISV sprouting (red arrows) is delayed and abnormal
at 25 hpf (e,f), but seemed to be recovering by 30 hpf (g,h). The views of embryos are lateral
with anterior left and dorsal up. Inserts c’ and d’ show transverse sections through the posterior
trunk. The number of embryos with notch5 or deltaC expression in the DA is given as a fraction
of the number of embryos analysed. Thirty-three hey2-morphant and thirteen wt fli1-egfp
transgenic embryos were analysed in (e–h). All morphants were injected with 5 ng hey2 MO.
REFERENCES
1. Lawson ND, Weinstein BM. In vivo imaging of embryonic vascular development using
transgenic zebrafish. Dev Biol 2002;248:307–318.