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Fig. 2 Exogenous Trh rescues the hatching phenotype of trh−/− zebrafish embryos. (A) Intracerebroventricular injection of intact trh−/− embryos. Brightfield images of a saline-injected embryo (top) and an embryo injected with Trh (bottom). Images on the right are magnified views of the boxed area in the corresponding left images that show HGCs (circled). The time elapsed from injection is shown at the top. (B) Quantification of hatching gland integrity in saline-injected (n = 8) and Trh-injected (n = 7) embryos. niGV, normalized inverted gray values. (C) Intramuscular injection of decapitated trh−/− embryos. Brightfield images of a saline-injected embryo (top) and an embryo injected with Trh (bottom). Images on the right are magnified views of the boxed area in the corresponding left images that show HGCs (circled). The time elapsed from injection is shown at the top. (D) Quantification of hatching gland integrity in saline-injected (n = 4) and Trh-injected (n = 5) decapitated embryos. (E) Confocal images showing colocalization of Trh receptors (green puncta) on HGCs in prehatching embryos. HGCs are outlined. (F) Effect of F0 deletion of Trh receptors on hatching success.