Additional Supporting Information may be found in the online version of this article.

JNR_22664_sm_suppinfoFig1.tif17564KSupporting Information Fig. 1. Identification of cell types in otocyst culture. Confocal micrographs showing expression of an epithelial marker, E-cadherin (red), and neuroblast marker, βIII tubulin (green) in a ventral otocyst preparation after 24 hr in culture. E-cadherin is concentrated in the epithelium (B,C,H,I) with very little staining associated with neuroblasts (B,C, E,F). βIII tubulin is highly expressed in neuroblasts (A,C,D,F) ans is expressed at much lower levels in the epihtelium (A,C,G, I). Scale bars = 100 μm. E, epithelium; N, neuroblasts
JNR_22664_sm_suppinfoFig2.tif3452KSupporting Information Fig. 2: Measurement of otic ganglion volume. The confocal micrographs show an example of an optical section through the cultured whole otocyst, highlighted by DAPI-stained nuclei, and the ganglion that formed during the incubation period, identified by the presence of NGN-1 GFP positive neuroblasts. The ganglion area measured for this section is highlighted with a dotted line. Equivalent measurements were made for each optical section and combined to calculate volume of the ganglion as described in Materials and Methods. O, otocyst; E, epithelium; G, ganglion. Scale bar = 100 μm

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