Fig. 7

Fig. 7

A Absence of dand5 repression in maternal zygotic (MZ) pkd2 mutant zebrafish at 10 somite stage (ss). B Quantification of dand5 asymmetry in controls (co) and pkd2 morphant (1–4 ng) zebrafish. Asymmetry was determined by picture analysis using ImageJ. Number (n) represents the number of analyzed specimens. Statistical analyses were done with one-sided Pearson’s chi-square test. C Animal cap luciferase reporter assay of full-length dand5.S 3′-UTR (cf. Figure 1A). The reporter construct was injected as mRNA either alone or in combination with high or low dose bicc1 mRNA, pkd2 mRNA or pkd2 TBMO. Gradual repression upon co-injection of high or low concentrations of bicc1 mRNA was observed. Administering only pkd2 mRNA or pkd2 TBMO (1 pmol) efficiently blocked or boosted luciferase expression, respectively. The data further indicate that in AC cells endogenous dand5 mRNA is post-transcriptionally regulated in a Pkd2-dependent manner. In the presence of a lower amount of bicc1 mRNA high-level, strong repression was achieved when pkd2 mRNA was co-injected, or further diminished upon knockdown of pkd2 using TBMO. N represents the number of independent experiments. A pool of 10 animal caps was analyzed per experiment and treatment. The results from reporter mRNA alone served as reference and were 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 (Bonferroni corrected) using the values of three individual experiments. p values, values for individual experiments, mean values, and standard deviations are found in the source data file. n.s. not significant, p < 0.05; ** highly significant p < 0.01; ***, very highly significant p < 0.001, RLU relative luciferase units, Lucluciferase.