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Fig. 4

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ZDB-IMAGE-180611-17
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Figures for Kim et al., 2017
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Fig. 4

Wdr11 knockdown in zebrafish causes defects associated with aberrant ciliogenesis

  • A, B Antisense morpholinos (MO) targeting either the exon 3–intron 3 (A) or exon 9–intron 9 (B) splice sites of zebrafish wdr11 result in microphthalmia (blue arrowheads), disorganized melanocyte migration (red arrowheads), a small cranium and reduced myotome formation, compared to control MO or uninjected embryos at 72 h post‐fertilization (hpf). Con MO, n = 303/306; wdr11 E3I3 MO, n = 157/178; wdr11 E9I9 MO, n = 94/105. Scale bar, 500 μm.

  • C. Birefringence analysis of wdr11 morphants compared to control embryos at 72 hpf reveals a requirement for wdr11 in muscle formation. Arrowheads indicate birefringence signal from skeletal muscle. Con MO, n = 40/40; wdr11 E3I3 MO, n = 20/20; wdr11 E9I9 MO, n = 20/20. Scale bar, 500 μm.

  • D. Ptc1 expression was reduced in the head (left panels) and myotome (right panel, arrowheads and asterisks) of wdr11 morphant embryos at 24 hpf, as shown by in situ hybridization (ISH). Con MO, n = 84/84; wdr11 E3I3 MO, n = 95/97; wdr11 E9I9 MO, n = 60/63. Scale bar, 100 μm.

  • E. Alcian blue staining of wdr11 E3I3 morphant embryos, compared to control at 120 hpf. Meckel's cartilage (mc), palatoquadrate (pq), ceratohyal (ch), ceratobranchials (cb) are indicated. Con MO, n = 86/87; wdr11 E3I3 MO, n = 124/128. Scale bar, 200 μm.

  • F. Sox10 ISH indicated a loss and reduction of neural crest cell (NCC) migration to OB (inset) and cranium (arrowheads), respectively, in wdr11 E3I3 morphants at 24 hpf. Con MO, n = 56/56; wdr11 E3I3 MO, n = 103/111. Scale bar, 200 μm.

  • G. Immunofluorescence staining of Gnrh (green) and acetylated tubulin (red) revealed reduced olfactory Gnrh neurons and ablation of cilia (arrowheads) in olfactory pits of E3I3 morphants at 72 hpf. Con MO, n = 20/20; wdr11 E3I3 MO, n = 18/20. Scale bar, 50 μm.

  • H. Acetylated tubulin staining (red) marking the cilia of control and E9I9 morphants demonstrates that knockdown of wdr11 results in reduced ciliogenesis in the olfactory pits (arrowheads). Con MO, n = 20/20; wdr11 E9I9 MO, n = 19/20. Scale bar, 20 μm.

  • I. Localization of Wdr11 (green) in the olfactory cilia (red, arrows) of WT embryos at 72 hpf. n = 10. Scale bar, 20 μm.

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