Figure 3

In vivo analysis of the role of the G4 present in pre-miR-150 during zebrafish embryo development. (a) Diagram of the strategy used to disrupt the formation of the G4 in the pre-miR-150 microinjected in zebrafish embryos. Reverse transcription followed by quantitative PCR (RT-qPCR) of miR-150 (b), myb (c), and miR-133a (d) using RNA samples from 24 hpf staged embryos microinjected with miR-150 or miR-133a precursors synthesized using either GTP (G-pre-miR-150 and G-pre-miR-133a) or 7-Deaza-GTP (7DG-pre-miR-150 and 7DG-pre-miR-133a). Ordinary 1-way ANOVA with Tukey multiple comparisons test, p < 0.005. Micrographs of representative 48 hpf staged larvae (lateral view, head to the left) microinjected with the RNA transcribed from pSP64T+dsRED plasmid containing no cloned pre-miRNA (control, (e)) and 7DG-pre-miR-150 (f), with the measured parameters indicated by blue, dashed lines in (e). Trunk length (g) and eye size (h) measurements of 48 hpf staged larvae microinjected with G-pre-miR-150 or 7DG-pre-miR-150. Ordinary 1-way ANOVA with Tukey multiple comparisons test, p < 0.001. Letters above each condition in the graphs represent statistical groups, where means with no letter in common are significantly different. For graphs shown in (b–d,g,h) p-values are detailed in Supplementary Table S7 and numerical values are detailed in Supplementary Table S8. Data shown represent results of one from three independent experiments.