(A) Live imaging of 30 hpf wild-type embryos treated with the indicated dose of U18666A starting at 4 hpf. U18666A-treated embryos exhibited a dose-dependent phenotype ranging from smaller otoliths at 24 μM to no otoliths, abnormal head, no circulating blood cells, and curved/twisted body axis at 96 μM. Arrowheads indicate the otic vesicle. Scale bars: 500 μm (whole embryo); 200 μm (otic vesicle). (B) Measurement of otolith diameters from DMSO- and U18666A-treated wild-type embryos at 30 hpf. n=118 (DMSO), 118 (24 μM), 114 (48 μM) and 94 (96 μM) (n=2 experiments). Two otoliths per embryo were measured. 0 indicates that otolith was not detectable. P<0.0001 for each treatment group compared with DMSO (Kruskal–Wallis test).
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