Fig. 5

Fig. 5

Skeletal and vascular phenotypes in rfc2 KO and rfc5 KO zebrafish. A: Defects in head cartilage formation in KO zebrafish at 5 dpf. Alcian blue staining, ventral view, anterior is to the left. B: Quantification of cartilage structures in rfc2 KO zebrafish. n = 14 for WT and n = 18 for KO. C: Defects in bone structures in KO zebrafish. Alizarin red S staining at 9 dpf. Lower panels are magnifications of upper panels, showing defects in bone and teeth development (arrows). n = 8 for rfc2 WT, n = 25 for rfc2 KO, n = 10 for rfc5 WT, and n = 23 for rfc5 KO. Ventral view, anterior is to the left. D: Vascular defects in KO zebrafish crossed with Tg(kdrl:gfp) at 7 dpf. Aortic arches are indicated by arrows. n = 9 for rfc2 WT, n = 27 for rfc2 KO, n = 14 for rfc5 WT, and n = 18 for rfc5 KO. Ventral view, anterior is to the left. E: Lateral view of aortic arches. Blood vessels were narrowed and hypo-branched in rfc2 KO, compared to WT and Het. F: Quantification of vessel diameter of the most posterior aortic arch (arrow in E) was measured using Image J software. n = 9 for WT, n = 18 for Het, and n = 9 for KO. Data are presented as mean ± SEM. Statistical significance was determined by the Kruskal–Wallis tests with post-hoc Dunn's multiple comparisons tests. ns, not significant, P > 0.05; ∗∗, P < 0.01; ∗∗∗, P < 0.001; ∗∗∗∗, P < 0.0001. Scale bar, 100 μm (A, C, D, E). dpf, days post fertilization; AW, articulation width; PL, Palatoquadrate length; M, Meckel's cartilage; m, maxilla; o, opercle; en, entopterygoid; br, branchiostegal ray 1; cb, ceratobranchial 5; WT, wild-type; KO, knockout; Het, heterozygous; SEM, standard error of the mean.