DEV042150 Supplementary Material

Files in this Data Supplement:

• Supplemental Table S1 -

• Supplemental Figure S1 -

  Fig. S1. Hedgehog-responsive genes are expressed in ventral jaw cartilages. (A) Ptc2 expression at 4 dpf. Strong expression can be seen in the ceratohyal and Meckel’s cartilage (black arrows), and in the branchial arches in the chondrocyte precursors (blue arrows). (B) Expression of the Hh transcription factors gli1, gli2 and gli3 in sibling and ptc2^−/− fish at 4 dpf. Both gli1 and gli3 appear upregulated in the mutants, compared with siblings, particularly in the cartilage precursors of the Meckel’s cartilage (red arrows) and ceratohyal (black arrow in gli3 pictures). By contrast, gli2 expression levels appear to be unchanged or slightly weaker in the mutants than in the siblings.

• Supplemental Figure S2 -

  Fig. S2. Only small changes to dermal bone formation are observed in Hedgehog signalling mutants. (A) Confocal stacks of wild-type and ihha^−/− opercula at 10 dpf, imaged with Tg(osx:nuGFP) (green) and Alazarin Red. (B) Brightfield and fluorescent images of wild-type and ptc1 4 dpf embryos. The arrowhead indicates osterix expression in the ectopic tissue between the eyes of the ptc1 mutant.

• Supplemental Figure S3 -

  Fig. S3. Premature ossification in ptc1 and ptc2 mutants is not accompanied by increased vascularisation. (A) Lateral views of head vasculature at 4 dpf; anterior to the right. In ptc1^−/−, the gill vasculature is reduced (white arrows), whereas there is no difference between ptc2^−/− and wild-type vasculature. (B) Confocal stack, taken in a wild-type embryo at 7 dpf, of the ceratohyal, showing flk-GFP (green), Alazarin Red (red) and merged image.