Rescue of enhanced angiogenesis in ptena-/-ptenb-/- mutants by exogenous ptena mRNA, LY294002 or sunitinib. The transgenic line, Tg(kdrl:eGFP), was used to visualize the vasculature at 4 dpf in wild-type (A,C,E,G) and ptena-/-ptenb-/- (B,D,F,H) embryos. Close-ups of the intersegmental vessel above the urogenital opening are shown in adjacent panels. (A–B2) ptena-/-ptenb-/- mutants display ectopic vessel growth compared with wild-type embryos. (C–D2) 10 pg synthetic ptena mRNA was injected at the one-cell stage into wild-type and ptena-/-ptenb-/- embryos. (E–F2) 25 μM LY294002 (LY) was applied from 70–72 hpf onwards. (G–H2) 20 μM sunitinib was applied from 70–72 hpf onwards. Images were taken using a confocal microscope with 20×. The numbers in the bottom right corner represent the total number of embryos treated/the number of embryos showing the phenotype depicted here. Anterior to the left, 20×, 2 μm step size.
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