EAF family genes act in multiple cellular responses including tumor suppression and embryonic development.In our previous studies,we found that Ear1/2 modulate convergence and extension(C & E) movements and pattern embryonic neuroectoderm during zebrafish embryos.Here,we reported that Eaf1/2 acted to limit TGF-β signals in germ layer formation.Knockdown Eaf1/2 in embryos caused expanded mesoderm and endoderm and led to the recovery of endoderm specification in mzoeptz257 mutant by limiting TGF-β signals.Gene expression profiling analyses in Eaf deleted or overexpressed mammalian cells convinced that roles of Eaf1 and Ear2 in inhibiting TGF-βsignals were conserved from fish to mammalian.By taking advantage of P53M214K mutant,TGF-β reporters and mechanisms detection,we revealed that Ear1 and Ear2 suppressed TGF-β transduction by synergistically suppressing non-P53 and P53-required TGF-β targets.P53 ectopic expression partially phenocopied ears morphants,P53M214K mutant exhibited subtle reduced endoderm and mesoderm,and knockdown of P53 in Ear1/2 absence cells could recovery expression of some TGF-β targets.