Gpc4 enhances the effects of full-length Wif1 in zebrafish embryos.

Approximately 2 nL of mRNA of a given concentration was injected into one cell stage embryos, and embryos were scored at 28–30 hours post-fertilization. Images show representative examples of the penetrance of the short-tailed phenotype compared to an uninjected control. In top panels dorsal is up and anterior is to the left. Bar graphs show percentage of short-tailed embryos. The data is pooled from two independent experiments; frequencies were scored and their percentages were averaged. See Materials and Methods for information on RNA preparation.

Wif1 expression in zebrafish embryos leads to decreased expression of a β-catenin-regulated reporter. (A, A′) DIC (A) and fluorescence (A′) images of embryos carrying the Tg(TOP:dGFP) reporter, showing GFP expression in the dorsal midbrain (arrow), anterior to the midbrain-hindbrain boundary. (B) Injection of 40 pg/nL of wif1 mRNA reduces GFP expression (arrow). (C) Injection of 5 pg/nL of kny mRNA does not change GFP expression. (D) GFP intensities were measured in living embryos and their statistically significant differences were verified by two-tailed Mann-Whitney U test. Statistically significant differences in GFP intensity were measured. No differences of statistical significance from were observed between wild-type and kny-injected embryos.

Acknowledgments
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