Figure Caption
Fig. 1
Disruption of mir430 transcription bodies does not substantially impact development.
a, Schematic representation of the maternal-to-zygotic transition in zebrafish embryos. b,c, Visualization of elongating RNAPII (Ser2P) with Fabs in WT embryos at the 256-cell (256c), 1,024-cell (1,024c), Oblong and Sphere stages (b) and in WT and mir430−/− embryos at the 128-cell stage (c). Shown are representative micrographs of individual nuclei, extracted from a spinning disk confocal microscopy timelapse. d, Schematic representation of a nucleus in WT, mir430−/− and mir430−/− with injected MiR430 embryos. e, Approach to assess rescue of miR430 activity, as previously described24. The expression of eGFP encoded by an mRNA with three perfect target sites for miR430 is compared with the expression of RFP encoded by an mRNA without such sites (Methods). f, Schematic representation of expected eGFP and RFP expression in embryos with active and inactive MiR430 microRNA activity. g, Representative micrographs showing eGFP and RFP expression in WT, mir430+/−, mir430−/− and mir430−/ with injected MiR430 embryos at 24 hpf. N = 3 biologically independent experiments. h, Rescue of miR430 activity was assessed in different genotypes at 24 hpf. Normalized eGFP signal in WT, mir430+/− and mir430−/− embryos without (left) and with (right) injected MiR430. N = 3 biologically independent experiments, n = 30 (WT without injected MiR430), n = 53 (mir430+/− without injected MiR430), n = 31 (mir430−/− without injected MiR430), n = 45 (WT with injected MiR430), n = 74 (mir430+/− with injected MiR430), n = 27 (mir430−/− with injected MiR430). Kruskal–Wallis tests were performed (without injected MiR430: χ2 = 67.892, d.f. = 2, P value = 1.809 × 10−15; with injected MiR430: χ2 = 2.5872, d.f. = 2, P value = 0.2743). When this test was statistically significant (P value < 0.05), pairwise comparisons with Bonferroni correction were performed using a pairwise Wilcoxon rank-sum test. A comparison was considered significant when adjusted P value was <0.05, and adjusted P values were reported using WT as reference. i, Rescue of miR430 activity as assessed by epiboly progression. Shown are representative micrographs of embryos at late epiboly stage in different genotypes. The misregulation of yolk internalization in mir430−/− embryos is indicated. j, Time at which epiboly is completed in different genotypes without (left) and with (right) injected MiR430 RNA. N = 3 (without injected MiR430) and N = 4 (with injected MiR430) biologically independent experiments, n = 19 (WT without injected MiR430), n = 34 (mir430+/− without injected MiR430), n = 19 (mir430−/− without injected MiR430), n = 27 (WT with injected MiR430), n = 53 (mir430+/− with injected MiR430), n = 28 (mir430−/− with injected MiR430). Kruskal–Wallis tests were performed (without injected MiR430: χ2 = 38.379, d.f. = 2, P value = 4.635 × 10−9; with injected MiR430: χ2 = 0.00080597, d.f. = 2, P value = 0.9996). When this test was statistically significant (P value < 0.05), pairwise comparisons with Bonferroni correction were performed using a pairwise Wilcoxon rank-sum test. A comparison was considered significant when adjusted P value was <0.05, and adjusted P values were reported using WT as reference. k, Time at which Kupffer’s vesicle appears in different genotypes without (left) and with (right) injected MiR430. N = 3 (without injected MiR430) and N = 4 (with injected MiR430) biologically independent experiments, n = 18 (WT without injected MiR430), n = 27 (mir430+/− without injected MiR430), n = 14 (mir430−/− without injected MiR430), n = 25 (WT with injected MiR430), n = 44 (mir430+/− with injected MiR430), n = 26 (mir430−/− with injected MiR430). Kruskal–Wallis tests were performed (without injected MiR430: χ2 = 26.038, d.f. = 2, P value = 2.218 × 10−6; with injected MiR430: χ2 = 2.543, d.f. = 2, P value = 0.2804). When this test was statistically significant (P value < 0.05), pairwise comparisons with Bonferroni correction were performed using a pairwise Wilcoxon rank-sum test. A comparison was considered significant when adjusted P value was <0.05, and adjusted P values were reported using WT as reference. l, Larvae at 48 hpf for different genotypes. Representative micrographs are shown. The malformation of trunk morphology and eye, the development of heart oedema, and the appearance of blisters at the tail tip in mir430−/− embryos are indicated (red arrowheads). Source numerical data are available in Source data.
Source data
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