Fig. 4

A Animal cap assay using a luciferase reporter mRNA which contained gdf3 3′-UTR sequences. Translation of gdf3 reporter was efficiently blocked by co-injecting bicc1 mRNA. N represents the number of independent experiments. A pool of 10 animal caps was analyzed per experiment and treatment. The result from reporter mRNA alone served as a reference and was set to 100% RLU. Relative values of single experiments are depicted as blue dots. Data of three experiments are presented as mean value (bar) ±standard deviation (error bar, SD). Statistical analyses were done with a one-sided Student’s t test for two independent means using the values of three individual experiments. Bgdf3 mRNA was not affected by bicc1 LoF. C Quantification of gdf3 expression in bicc1 morphants at the LRO margin. Dgdf3 GoF rescues nodal1 expression in bicc1 morphants. Representative nodal1 staining in left sLRO cells is shown for control (co), bicc1 morphant, and rescued specimens. E Quantification of the bicc1 MO rescue of nodal1 expression by gdf3. F, G Left-asymmetric pitx2 expression (arrowhead) is restored in bicc1 morphants by co-injecting gdf3 mRNA. MO pmol/embryo: bicc1 SBMO (L and S, each 1). Asterisks in B and D mark injected side. Numbers (n) in C, E, and G represent analyzed specimens from more than independent experiments. Statistical analyses were done with one-sided Pearson’s chi-square test, which was adjusted for multiple comparisons by Bonferroni–Holm (C, E) or Bonferroni (G). n.s. not significant, p > 0.05; ** highly significant p < 0.01; ***, very highly significant p < 0.001. p-values, mean values, SD and listing of individual experiments can be found in the source data file. Scale bars in B and D represent 100 µm and in F 1 mm. RLU relative luciferase units, st. stage, a anterior, l left, r right, p posterior, d dorsal, v ventral.