Knockdown of nme3 in zebrafish results in cyst formation and left-right asymmetry defects. A, zebrafish embryos injected with control (CTL) MO, nme3 SB MO, or nme3 TB MO at 48 hpf. In contrast to the control embryos, nme3 morphants showed a curly-tail phenotype. Scale bars, 100 μm. Nme3-depleted zebrafish embryos showed pronephric cyst formation, visualized using the transgenic Tg(wt1b:EGFP) line (white asterisks). Scale bars, 50 μm. Histological sections of nme3 morphants stained by hematoxylin and eosin confirmed cyst formation (black asterisks). Scale bars, 10 μm. B, quantification of the percentage of cyst formation caused by knockdown and CRISPR-mediated mosaic indel formation of nme3 at 48 hpf (*, p < 0.05; ***, p < 0.001; t test; error bars represent S.D.). C, representative pictures of 18-somite-stage zebrafish embryos after WISH for spaw with normal, bilateral, or inverted spaw expression (black arrows). D, quantification of the percentage of altered spaw expression in nme3 zebrafish morphants. E, examples of zebrafish embryos with normal (left) and reversed (middle and right) heart looping, visualized by WISH for the heart-looping marker cmlc2 at 48 hpf. The red arrows point to the atria. F, quantification of the percentage of heart laterality defects caused by nme3 depletion in zebrafish embryos at 48 hpf.
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