Fig. 2

Elevated N-fucosylation dorsalizes zebrafish embryos. (A) Variable degrees of dorsalization in embryos over-expressing mSlc35c1. (B) Quantification of the severity and penetrance of dorsalization in each treatment. (C and D) qPCR of the ventral transcriptional factors vox and vent in WT, GDP-Fuc injected or mSlc35c1 expressing embryos. (E–L) in situ hybridization with dorsal (chordin and gsc) and ventral (eve1) specific markers at shield stage (6hpf). (E–H) chordin expression in treated embryos (I–L) eve1 and gsc expression. The expression angles of chordin and gsc of embryos with each treatment are summarized in (M) and (N) respectably. The sample sizes for each treatment are as follows: wt (N=15), GDP-Fuc(N=13), mSlc35c1(N=18) and mSlc35c1 plus GDP-Fuc (N=21) for (M); and wt (N=19), GDP-Fuc (N=15), mSlc35c1(N=21) and mSlc35c1 plus GDP-Fuc (N=22) for (N). For chordin angle, ordinary One-way ANOVA between sample group means, p=0.0002. For gsc angle, ordinary One-way ANOVA between sample group means, p=0.0001. Note: ^** Tukey′s procedure after ANOVA p<0.01. Null hypothesis for Χ² between embryos injected with 1 ng mSlc35c1 and with GDP-Fuc plus 1 ng mSlc35c1 p=1.41E-04; for Χ² between embryos injected with 2 ng mSlc35c1 and with GDP-Fuc plus 2 ng mSlc35c1 p=8.99E-03.