Effects of KN-93 on embryonic development. Gross morphology was assessed at 72 hpf in untreated embryos (A) and in embryos continuously treated from cleavage stage with 2.5 μM KN-93 (B), 5 μM KN-93 (C) and 10 μM KN-93 (D). CaMK-II kinase activity assays were conducted on 3 dpf whole embryo lysates and 3dpf partially purified CaMK-II in the presence of increasing concentrations of KN-93 (E). Heart rates at 72 hpf were averaged from at least 20 embryos per drug concentration (F). 10 μM KN-93 treatment resulted in otolith defects at 30 hpf (G, H) and 72 hpf (I, J). Scale bar (50 μm) in G applies to G, H and in I applies to I, J. Otolith defects assessed at 72 hpf were tabulated for 125 control and 182 10 μM KN-93 treated embryos (K). P-CaMK-II (red) was diminished in 10 μM KN-93 (M) treated embryos compared to control embryos (L, arrowhead) at 24 hpf but gata3 was expressed in the otic vesicles of both control (N) and 10 μM KN-93 (O) treated 24 hpf embryos. Scale bars=5 μm (L, M) and 50 μm (N, O).
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