Fig. 3. Phenotypes associated with lack of both bon and sur activity. Wild-type (A1-4), bon (B1-4), bonMO (C1-4), MZsur (D1-4), MZsur;bon (E1-4) and MZsur;bonMO (F1-4) embryos at 30 hpf. All images are lateral views except A3-F3, which are ventral views. (A1-F1) Comparison of embryos in a lateral view. The bon (B1) and bonMO (C1) embryos have pericardial edema (arrowhead) and enlarged yolk extension (arrow). MZsur;bon (E1) and MZsur;bonMO (F1) embryos have an anterior bulb-like structure (arrowhead; also arrow in E3 and F3). (A2-F2) Lateral view of head region, showing the absence of the hatching gland (arrowhead) in MZsur;bon (E2) and MZsur;bonMO (F2) embryos but present in others. (A3-F3) Ventral view of head region, presence of two hearts (cardiac bifida; arrowheads), in bon (B3) and bonMO (C3), single heart in MZsur (arrowhead in D3) and no heart in MZsur;bon (E3) and MZsur;bonMO (F3) embryos. (A4-F4) Lateral view of trunk and tail region, all embryos display normal notochords (arrow). MZsur;bon and MZsur;bonMO embryos exhibit accumulated blood (arrowhead), owing to defects in circulation. 100% of MZsur;bon embryos lacked hatching gland, heart and ventral CNS (n=41). By contrast, 81% of MZsur embryos had a heart, hatching gland and two eyes, 15% had a heart and hatching gland and fused eyes, and 3% had a heart, no hatching gland and fused eyes (n=85).

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