Figure 1—figure supplement 2.

Figure 1—figure supplement 2.

(A–D) Dorsal views of 102 hpf WT (A), vegfab^-/- (B), vegfc^-/- (C), and vegfd^-/- (D) cranial vasculature visualized by Tg(kdrl:EGFP) expression. A significant fraction of vegfab^-/- and vegfc^-/- larvae lacked the DLV at 102 hpf. (E and F) Percentage of the fish of indicated genotype with and without the DLV at 102 (E) and 154 (F) hpf. A significantly higher fraction of vegfab^-/- and vegfc^-/- fish lacked the DLV at 102 hpf (47% and 40%, respectively) than WT (0%) (n = 13 for WT, n = 19 for vegfab^-/-, n = 25 for vegfc^-/-, and n = 19 for vegfd^-/- fish examined at 102 hpf). At 154 hpf, while 63% of vegfab^-/- fish exhibited the ‘No DLV’ phenotype, only 13% of vegfc^-/- fish showed this phenotype (n = 16 for WT, n = 27 for vegfab^-/-, n = 23 for vegfc^-/-, and n = 26 for vegfd^-/- fish examined at 154 hpf). Only a small fraction of vegfd^-/- fish examined lacked the DLV at 102 and 154 hpf (5% and 0%, respectively). (G–H") Dorsal views of 96 hpf vegfab^+/+ (G–G") and vegfab^-/- (H–H") heads of larvae that carry the Tg(kdrl:EGFP) transgene (G and H) and that were injected with streptavidin-conjugated Qdot 655 nanocrystals (G’ and H’) (merged images shown in G" and H"). In vegfab^+/+ fish (n = 10), the DLV is visualized by both Tg(kdrl:EGFP) expression (G) and blood-circulating Qdot 655 nanocrystals (G’). In contrast, in the vegfab^-/- larvae that lack the Tg(kdrl:EGFP)-positive DLV (H, n = 8), blood-circulating Qdot 655 nanocrystals did not label the DLV (H’), showing the lack of a perfused, functional DLV. Yellow arrows point to the presence and absence of the DLV in vegfab^+/+ and vegfab^-/- larvae, respectively. Scale bars: 50 µm.