Zebrafish borcs8 F0 knockout larvae exhibit developmental defects. (A) Morphology of zebrafish wild-type (WT), Cas9 control and borcs8 F0 knockout (KO) larvae at 3 days post-fertilization (dpf). Scale bars = 1 mm. (B) Frequency of phenotypes observed for wild-type, Cas9 control, borcs8 F0 KO and human BORCS8 mRNA rescue (Rescue) larvae (N = 4, n = 32–72). (C–E) Body length (C), head size (D) and eye size (E) of wild-type, Cas9 control, borcs8 F0 KO and Rescue larvae at 3 dpf (N = 3, n = 8–9 for head size and body length, n = 16–18 for eye size). (F) Haematoxylin and eosin staining of paraffin-embedded brain sections from the midbrain of 5 dpf wild-type and borcs8 F0 KO larvae. Scale bar = 100 μm. (G) Quantification of brain size of borcs8 F0 KO larvae (N = 9) relative to wild-type (N = 15) and Rescue (N = 4) at 5 dpf. (H) Phalloidin staining of muscles in borcs8 F0 KO and wild-type larvae at 3 dpf. (I and J) Comparisons of dorsal or ventral myotome area (I) and myoseptum angle (J) between wild-type (N = 3, n = 8) and borcs8 F0 KO larvae (N = 3, n = 8–9). All data are represented as the mean ± standard error of the mean (SEM). Statistical significance was calculated by one-way ANOVA followed by Tukey’s multiple comparisons tests, or Student’s t-test. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. N = replicates; n = sample size; ns = not significant.
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