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man1a2 knockdown results in laterality defects in zebrafish. (A) Whole-mount in situ hybridization images showing myl7 (cardiomyocytes) and foxa3 (endodermal cells) expression at 36 hpf, which was used to determine the laterality of embryos. Three patterns of heart looping (normal, reversed, and midline) and two patterns of visceral organ position (normal and reversed) were detected and were quantified in graphs. Numbers in the graph indicate the number of larvae in each group. Arrows and arrowheads point to the liver and the dorsal pancreas, respectively; dotted lines outline the heart. Scale bars: 100 μm. (B) Confocal images of Kupffer’s vesicle showing the expression of dusp6:d2GFP (green, Kupffer’s vesicle cells) and acetylated tubulin (red, cilia). Both cilia length and number were significantly reduced in man1a2 MO-injected larvae (n = 11) compared to controls (n = 9), as shown in graphs. Scale bar: 50 μm. Error bars: ± SEM. *p < 0.001; **p < 0.005.
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