Figure 3.

Localization of Pcdh15a in mechanotransduction mutants. Immunolabel of Pcdh15a (magenta) and phalloidin-labeled actin (green) in a wild-type sibling (A, A′) and lhfpl5a^tm290d mutant (B, B′). An example of remaining signal in a mutant hair bundle is shown (arrowhead). C–F′′′, images of stably expressed EGFP-tagged Pcdh15a-CD3 in mechanotransduction mutants at 6 dpf. Representative confocal images are shown. The white arrowheads and brackets indicate mature hair bundles, whereas the yellow arrowheads and brackets indicate immature hair bundles at the periphery of the cristae. C–C′′′, lhfpl5a^tm290d mutant; Pcdh15a-CD3-EGFP localized predominantly to the soma, but faint signal was occasionally detectable at the tips of the longest stereocilia (inset; scale bars 2 μm). D–D′′′, cdh23^nl9 mutant; Pcdh15a-CD3-EGFP is absent from hair bundles in mature hair cells, but present in immature bundles. E–E′′′, myo7aa^ty220d mutant; Pcdh15a-CD3-EGFP was absent in hair bundles of mature cells, similar to cdh23 mutants. F–F′′′, myo6b^tn3137 mutant; Pcdh15a-CD3-EGFP localized at the tips of hair bundles in a staircase pattern (arrowheads in F′′′). Black bracket in F′ indicates a fused bundle phenotype. C′′′, D′′′, E′′′, F′′′, Higher-magnification image of area from C–F, respectively (outlined with yellow box). n > 5 larvae for each genotype. Scale bars in C–C′′, D–D′′, E–E′′, F–F′′, 10 μm. Scale bars in A–B′, C′′′, D′′′, E′′′, F′′′, 5 μm.