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Fig. 1 Zebrafish embryos carrying a loss-of-function mutation in the trh gene cannot hatch. (A) Annotated illustration of the zebrafish trh gene and mutation strategy. Guide RNA was designed to target the translation start site and generated a 4-bp deletion immediately downstream of the start codon. (B) Hatching percentage in clutches resulting from crosses of different genotypes. Bottom images show representative pictures of the clutches at 3 dpf. n = 300 eggs per cross. (C) Brightfield images of trh+/+ (wild-type), trh+/−, and trh−/− mutants at 3 dpf. White arrowheads mark the hatching gland. The insets show magnified views of the hatching glands.