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Fig. 2 Embryonic development and growth abnormalities in ube3a mutants. (a) Representative images of zebrafish embryos from 4 h post-fertilization (hpf) to 8 hpf, showing the progression of epiboly in WT and mutant embryos. (b) Quantitative analysis of blastoderm thickness at 5 hpf (N = 11 for WT, N = 9 for mutant), and percentage of epiboly progression at 6 hpf (N = 11 for WT, N = 11 for mutant), 7 hpf (N = 11 for WT, N = 10 for mutant), and 8 hpf (N = 11 for WT, N = 10 for mutant). (c) Hatching rate between WT and mutants each 2 h from 48 hpf to 60 hpf. (d) Representative cardiac/yolk-sac edema in 4 dpf ube3a mutant embryos. Note that the severe cardiac/yolk-sac defects were observed in ube3a mutants, and at low frequency in WT clutches (see Fig. 2e), including AB strain, and were typically non-viable Left: cardiac/yolk-sac edema; right: cardiac/yolk-sac edema and body curvature. (e) Percentage of developmental abnormalities observed between 2–5 dpf. (f) Survival rate of WT and mutant zebrafish. (g) Interorbital distance at 4 dpf in WT (N = 31) and mutants (N = 23), representative images (left) and quantification (right). (h) Distance between the optic tecta, in WT (N = 7) and mutants (N = 11), representative images (left) and quantification (right). (i) Body length at 4 dpf, in WT (N = 24) and mutants (N = 22), representative images (left) and quantification (right). The data are presented as the mean ± SEM. ns: non-significant, *P < 0.05, **P < 0.01 and ***P < 0.001.