Fig. S4

Additional data related to Figure 6 (A) Quantification of artery hyperbranching in wildtype (n=20), vegfc^hu5055 heterozygous (n=43) and homozygous (n=23) embryos injected with a flt1 morpholino (MO). There are significantly fewer hyperbranched aISVs in vegfc heterozygous and mutant embryos when compared with vegfc wildtype embryos. (B) Sequence details for Talen induced mutagenesis. Talen induced mutagenesis by injection of forward and reverse Talens targeting the red and blue regions of the dll4 locus. Mutagenesis induced a 2bp deletion in the exon2 of the dll4 coding sequence (dll4uq10bh), predicted to generate a null allele due to the introduction of a stop codon at amino acid 47. (C) Confocal images of the trunk of 5dpf Tg(flt11:yfp);Tg(-5.2lyve1b:dsRed) zebrafish embryos showing hyperbranching (arrows) of aISVs in dll4uq10bh heterozygous embryos, which is absent in wildtype embryos. (D) The hyperbranching of aISVs at 3dpf following injection of flt1 MO (n=128) is partially rescued by co-injection with either kdr (n=120) or flt4 MO (n=91). For above analysis, error bars represent mean +/- sem; ****p<0.0001, ***p<0.001, ** p<0.01, *p<0.05 from one way ANOVA.