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Fig. 2 Zebrafish ndufaf2 morpholino–injected mutants exhibit a ciliopathy phenotype. (A) Protein levels of control morpholino (MO)–injected, ndufaf2 morpholino–injected, and p53 morpholino–injected embryos and ndufaf2 morpholino–injected embryos re-expressing NDUFAF2WT. Western blot analysis was performed with antibodies against NDUFAF2 and GAPDH. (B) Transmitted light images of body shape in control MO–, ndufaf2 morpholino–, and p53 morpholino–injected zebrafish larvae and ndufaf2 morpholino–injected zebrafish larvae re-expressing NDUFAF2WT, at 5 days post-fertilization (dpf). (C) Quantification of body curve in control MO–, ndufaf2 morpholino–, and p53 morpholino–injected zebrafish larvae and ndufaf2 morpholino–injected zebrafish larvae re-expressing NDUFAF2WT, at 5 dpf; >30 larvae analyzed for each independent experiment. (D) Transmitted light images of heart in control MO–, ndufaf2 morpholino–, and p53 morpholino–injected zebrafish larvae and ndufaf2 morpholino–injected zebrafish larvae re-expressing NDUFAF2WT, at 5 dpf. Arrows indicate the heart. (E) Quantification of heart failure in control MO–, ndufaf2 morpholino–, and p53 morpholino–injected zebrafish larvae and ndufaf2 morpholino–injected zebrafish larvae re-expressing NDUFAF2WT, at 5 dpf; >30 larvae analyzed for each independent experiment. (F) Confocal images of Kupffer’s vesicle (KV) cells labeled with cilia markers, Arl13b (green) and pGlu-Tu (red), in control MO–, ndufaf2 morpholino–, and p53 morpholino–injected zebrafish larvae and ndufaf2 morpholino–injected zebrafish larvae re-expressing NDUFAF2WT, at 8 somite stage. Scale bar: 20 μm. (G) Quantification of KV cilia number in control MO–, ndufaf2 morpholino–, and p53 morpholino–injected zebrafish larvae and ndufaf2 morpholino–injected zebrafish larvae re-expressing NDUFAF2WT; >30 larvae analyzed for each independent experiment. (H) TEM images of ndufaf2-mutant zebrafish showing shortened and disorganized photoreceptor outer segments at 5 dpf. Representative images of control MO– and ndufaf2 morpholino–injected zebrafish larvae and ndufaf2 morpholino–injected zebrafish larvae re-expressing NDUFAF2WT, at 5 dpf. Boxes show areas of enlarged mitochondria. M, mitochondria; M*, swollen mitochondria; OS*, shorter outer segment of photoreceptor. Scale bars: 2 μm. The bars in each graph represent mean ± SD. Exact P values are indicated. ANOVA followed by Tukey-Kramer multiple-comparison test.