Fig. 1

Morphology of the scube mutant family at 3 dpf. (A) Genetic features of scube1 mutants. The scube1 gene comprises 23 exons and encodes a protein of 994 amino acids. ZFN editing targeted the second exon, producing a 13-base pair deletion. This mutation introduces a premature stop codon at position 110. The mutated Scube1 protein is predicted to contain only one EGF domain and no other domains. (B) Genetic features of scube3 mutants. The scube3 gene consists of 22 exons and encodes a protein of 1002 amino acids. ZFN editing targeted exon 17, resulting in a 4-base pair deletion that generates a premature stop codon at position 850. The truncated Scube3 protein is predicted to lack the CUB domain. (C) Morphology of scube mutants at 3 dpf, sample size n = 3–10 embryos. Wild-type larvae at 3 dpf display a straight body axis. Similarly, scube1, scube3, and double mutants (scube1−/−scube3−/−) also maintain a straight body morphology. In contrast, scube2−/−, scube1−/−scube2−/−, scube2−/−scube3−/−, and triple mutants (scube1−/−scube2−/−scube3−/−) exhibit a curved body phenotype, characteristic of Hh signalling disruption. This curved body morphology resembles phenotypes caused by treatment with Cyclopamine, an inhibitor of the Hh pathway.