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Fig. 2 Eafs antagonize Wnt/β-catenin signaling in anterior neuroectoderm patterning. (A) eaf1 and eaf2 counteract wnt8a-mediated forebrain defects. (a) In situ hybridization for the anterior neuroectoderm marker opl in wild type (WT) and in the presence of wnt8a mRNA alone or together with eaf1 or eaf2 mRNA. (b) Scoring of opl expression: black, reduced; white, normal; gray, expanded. (B) eaf1 or eaf2 partially counteracts the strong reduction in opl expression that occurs after exposure to the Gsk3 inhibitor BIO, which enhances Wnt/β-catenin signaling. opl expression is scored as in A. (C) eaf1 and eaf2 rescue forebrain defects in apc mutants. opl expression is scored as: black, strongly reduced; blue, reduced; white, normal; gray, increased. (D) Heat shock-induced expression of dkk1 at 50% epiboly rescues forebrain defects in Eaf morphants. opl expression is scored as in A. (E) dn-Tcf partially rescues the anterior brain defects in Eaf morphants. Embryos were scored by six3b expression: black, reduced; white, normal; gray, increased. Aa,Ba,Ea, dorsal views, anterior to the left; Ca,Da, lateral views, anterior to the left.