Effects of eaf1 and eaf2 knockdown and overexpression on zebrafish embryos. (A) Morphology of representative embryos injected with the indicated MOs (8 ng/embryo) at 30 hpf. STD-MO, standard MO. (B) (a-f) In situ hybridization for the anterior brain marker six3b in the presence of the indicated MOs. (g) Scoring of six3b expression: black, reduced expression; white, normal expression. (C) (a-c) Morphology of representative embryos injected with the indicated mRNAs (200-500 pg/embryo). zeaf1 and zeaf2 refer to zebrafish eaf1 and eaf2. (d) Morphology was scored as normal (white) or severely defective (black) in eaf1- and eaf2-overexpression embryos. The expanded dorsal part is indicated by the black line. (D) eaf1 and eaf2 function as forebrain inducers in embryos. Embryos injected with eaf1 and eaf2 mRNA displayed increased expression of six3b (a,b) and opl (g,h), similar to embryos injected with dn-Tcf (d,j) or frzb (e,k) mRNA, but in contrast to embryos injected with wnt8a mRNA (f,l); egr2b exhibited reduced expression in embryos injected with eaf1 and eaf2 mRNA (m,n), similar to embryos injected with dn-Tcf (p) or frzb (q) mRNA. Ba-f,Da-q, dorsal views, anterior to the left.
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