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Fig. 3 Harmonin is required in otic sensory patches for normal bundle development. (A–C) Phalloidin-stained F-actin (green) and anti-zebrafish harmonin antibody labeling (red) in 80-hpf wild-type ears show localization of harmonin relative to the stereocilia in the five otic sensory patches (ac, anterior crista; am, anterior macula; lc, lateral crista; pc, posterior crista; pm, posterior macula) and in the neuromasts (n). The anti-harmonin antibody signal is reduced in sensory patches and in neuromasts in ush1c mutants (B) and ush1c-MO-injected larvae (C). (D–L) High-magnification view of phalloidin-stained hair bundles in the anterior and posterior maculae of 5 dpf larvae. Hair bundle number is reduced and organization of the existing bundles is disrupted in mutant (G–I) and morphant (J–L) anterior (G,J) and posterior (H,K) maculae, compared with the anterior and posterior maculae of wild-type controls (D and E, respectively). Closeup images of hair bundles of the anterior maculae (F,I,L) show bent and splayed bundles in mutants and morphants. For mutant analysis, anterior macular wild-type average=54.6±3.5 bundles; n=8; ush1c mutant average=33±3.5; n=7; P<0.0001. Posterior macula wild-type average=68.2±2.6 bundles; n=9; ush1c mutant average=40.5±4.2; n=10; P<0.0001. For the morpholino experiment, anterior macula control average=41.8±4.8 bundles; MO average=21.8±3.8; n=5 for each group; P<0.0001. Posterior macula control average=71.8±4.6 bundles; MO average=48.2±6.9; n=5 for each; P<0.0001, 5 dpf. Scale bars: 50 μm (A–C); 10 μm (D–L).