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

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DVDY21836SuppMoviesS1A1.avi494KVideos with 3D projections of <I>osx</I>:mCherry expression in branchiostegal rays, operculum, precaudal vertebrae, and caudal fin, respectively, corresponding to Figure 2K–M made by laser scanning microscopy are available from the authors on request. Supp. Fig. S1A1–C2. Movies of life imaging analysis of <I>osx</I>:mCherry expression in the parasphenoid (head), anterior vertebrae and caudal fin, respectively, corresponding to Figures 4A–C. <B>S1A1, B1, C1:</B> <I>osx</I>:mCherry expression only. <B>S1A2, B2, C2:</B> Overlays of mCherry images with images from bright field and GFP filter settings for (S1A2) and bright field for (S1B2, C2). Supp Movie S1A1.
DVDY21836SuppMoviesS1A2.avi2072KSupp Movie S1A2.
DVDY21836SuppMoviesS1B1.avi753KSupp Movie S1B1.
DVDY21836SuppMoviesS1B2.avi6704KSupp Movie S1B2.
DVDY21836SuppMoviesS1C1.avi408KSupp Movie S1C1.
DVDY21836SuppMoviesS1C2.avi4935KSupp Movie S1C2.
DVDY21836SuppFigsS2.tif763KSupp. Fig. S2. <B>A,B:</B> Ventral view of the head showing <I>osterix</I> expression at 10 days postfertilization (dpf; A) and double staining for cartilage (Alcian blue) and bone (Alizarin red) at 20 dpf (B), anterior to the left. Arrow and arrowheads mark <I>osterix</I> expression (A) and ossification centers (B) surrounding the basihyoid and ceratohyal, respectively.
DVDY21836SuppFigsS3.tif1315KSupp. Fig. S3. <I>osx</I>:mCherry-<I>osc</I>:eGFP double transgenic medaka. <B>A,B:</B> Ventrolateral view of the head showing expression of <I>osx</I>:mCherry (A) and <I>osc</I>:eGFP (B) in branchiostegal rays (bs) and operculum (op) at 1 days post hatching (dph). Note cells expressing both transgenes (arrowhead) and cells expressing osx:mCherry only (arrows). <B>C:</B> Overlay of (A) and (B). e, eye.
DVDY21836SuppFigsS4.tif1903KSupp. Fig. S4. Colocalization of transgenic mCherry and endogenous <I>osterix</I> expression in a medaka embryo at 5 days postfertilization (dpf) as analyzed by whole-mount RNA in situ hybridization. <B>A:</B> Ventral view of the head showing mCherry transcripts in red. <B>B:</B> Ventral view of the same embryo as in A after second staining for <I>osterix</I> transcripts (blue). Boxes in A and B represent higher magnification views of the operculum (op) and the first branchiostegal ray (bs), respectively.

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