DEV024034 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. 3D projections of micro CT scans of a vertebra of a 3-month-old wild type. The centrum exhibits a central opening.

• Supplemental Figure S2 -

  Fig. S2. 3D projections of micro CT scans of vertebrae of a 2-month-old stocksteif juvenile. One of two mutant centra still exhibits a central opening, while the central canal from the second vertebra is completely filled with mineralized tissue.

• Supplemental Figure S3 -

  Fig. S3. Osterix expression in siblings versus mutants. Expression of osx is not changed in 4 dpf siblings (A,B) versus mutants (C,D). Operculum is encircled in B,D. 5ba, 5th branchial arch; ec, ectopterygoid; en, entopterygoid; op, operculum; ps, parasphenoid.

• Supplemental Figure S4 -

  Fig. S4. Cyp26b1 expression in zebrafish. (A-E) Cyp26b1 expression in zebrafish at 3 dpf. (F-I) Cyp26b1 expression in zebrafish at 6 dpf. For sectioned material, in situ hybridization was performed on whole-mount embryos, and subsequently sections were cut using a vibratome (100 µm; B,G) or a microtome (10 µm; C,D). Expression of cyp26b1 is seen in the hindbrain, branchial arches, pectoral fins and ossified structures such as the operculum (encircled in E and H) and the parasphenoid (F,G). ba, branchial arches; br, brain; cl, cleithrum; ep, ethmoid plate; mc, meckel’s cartilage; no, notochord; oc, orbital cartilage; pf, pectoral fin; ps, parasphenoid. (F,G) Embryos that were only shortly stained show extensive staining in the parasphenoid. (H,I) Sibling (H) and sst^sa0002 mutant (I) embryos showing massive upregulation of cyp26b1 expression in mutants. Identically treated sibling embryos are depicted. Genotypes were determined by sequence analysis. Other than the embryo in I, all embryos are wild type.