Fig. 4

Establishment of gon4lb-null zebrafish and rescue using human GON4L mRNA.a Target design to knockout gon4lb using CRISPR/Cas9. A crRNA targeted exon 2 of gon4lb and resulted in a 13 bp deletion that generated a premature termination codon to knockout. b Representative images of 50-hpf zebrafish embryos from the gon4lb-knockout line: gon4lb+/+, gon4lb+/?, gon4lb?/?, and gon4lb?/? injected with human GON4L mRNA (rescue). All images are lateral views, with the anterior surface to the left. Scale bar: 200 µm. c Representative images of eye size and head size of 50-hpf zebrafish embryos from the gon4lb-knockout line. All images are dorsal views, with the anterior aspect at the top. The blue line delineates the contour of the head, and the pink line delineates the contour of the eye. Scale bar: 50 µm. Quantitative data showing eye (d) and head size (e) of 50-hpf zebrafish embryos: gon4lb+/+ (n = 12), gon4lb+/? (n = 12), gon4lb?/? (n = 10), and gon4lb?/? with rescue (n = 15). f Representative images of the body length of 50-hpf zebrafish embryos. All images are dorsal views, with the anterior aspect at the top. The red line indicates the position for measuring the body axis. Scale bar: 200 µm. g Quantitative data showing the body length of 50-hpf zebrafish embryos. Sample numbers are as follows: gon4lb+/+ (n = 30), gon4lb+/? (n = 30), gon4lb?/? (n = 31), and gon4lb?/? embryos with rescue (n = 17). Data are shown as the mean ± SEM; ****p ? 0.0001, §§§§p ? 0.0001 using one-way ANOVA with post hoc Tukey?s test.