Fig. 4

A–D Immunocytochemical labeling of primary cilia with an anti-acetylated tubulin antibody within Kupffer’s vesicle in uninjected (A), mak morpholino-injected (B), and morpholino-injected zebrafish that simultaneously received canonical MAK^CI (D) or retinal MAK^RI mRNA (D). E Quantification of mean cilia length measured in each treatment group shown in A–D. MAK morphants displayed significantly longer cilia compared to uninjected siblings (uninjected vs MO only; p = 0.0004). Sequential injection of retinal MAK^RI mRNA or canonical MAK^CI mRNA significantly shortened primary cilia length (MO only vs MO + RNA; p < 0.0001 for both). F–H Light micrographs of wildtype (F), MAK morpholino-injected (G), and MAK morpholino and retinal MAK mRNA injected (H) demonstrating normal overall morphology among groups. I Histogram depicting the number of responses in the vision startle assay of wild-type, MAK morpholino-injected (MAK Mutant) and MAK morpholino/MAK^RI mRNA injected (Mutant + MAK^RI mRNA). MAK morphants had a significant decrease in average number of responses compared to wild-type fish (Wildtype vs MAK Mutant; p < 0.05). Injection of retinal MAK mRNA into MAK mutants partially rescued visual responses (i.e. no significant difference between wild-type and treatment groups). Kruskal-Wallis test with Dunn’s multiple comparisons. Scale bars in A-D = 10 μm.