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Fig. 6-S1
Artificial, uniform Nodal inhibition rescues morphology at 1 dpf in lefty1-/-;lefty2-/- mutants.
Wild type and lft1-/-;lft2-/- mutant embryos were exposed to 0, 1.25, 2.5, 3.75, 5, or 6.25 ?M SB-505124 (Nodal inhibitor drug) starting at the 8 cell stage, sphere stage, or shield stage. Drug was removed at 24 hr post-fertilization. Rescue at earlier stages requires lower concentrations (2.5?3.75 ?M) of SB-505124 than rescue at later stages (5?6.25 ?M). (A?B) Unexposed wild type (A) and lft1-/-;lft2-/- mutant (B) embryos at 1 dpf. (C?Q?) Embryos exposed to SB-505124 from the 8 cell stage to 24 hpf (C?G?), sphere stage to 24 hpf (H?L?), and shield stage to 24 hpf (M?Q?) at 1 dpf. Number of embryos with same phenotype as image/total is indicated in bottom right of images. Cyclopia and loss of head and trunk mesendoderm are typical Nodal loss-of-function phenotypes (Gritsman et al., 1999; Hagos and Dougan, 2007), and can be observed in both wild type and mutant embryos treated with higher amounts of SB-505124 (e.g., F and F?). Wild type embryos exposed to 3.75 ?M at the 8 cell stage (E) exhibited cyclopia (15/27, shown here) and normal morphology (12/27). Wild type embryos exposed to 3.75 ?M at sphere stage (J) exhibited mild cyclopia (6/30), moderate cyclopia (13/30, shown here), necrosis and cyclopia (1/30), normal morphology (9/30), and 1/30 died. Some images shown here are also used in Figure 6.