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Supporting Figure 3 Craniofacial phenotypes induced by wildtype xduct and ductin-independent gain-of-function reagents. Ductin loss-of-function produced a range of craniofacial phenotypes (see Fig. 2). Reagents predicted to act as gain-of-function agents, including the sodium-hydrogen exchanger nhe3, and a yeast proton pump pma1.2, were also tested for their affects on morphology. We also examined the effect of wildtype ductin mRNA (xduct). Tadpoles were scored and imaged between stage 45 and 48. All three of these constructs produced a similar suite of phenotypes to each other and to xduct-noTM4 and concanamycin. In all images, the construct injected is indicated and arrowheads point to abnormal structures. All views are dorsal except where indicated by a V (ventral) or a P (profile). Anterior is up except in I, where anterior is left and dorsal is up. A: Tadpole illustrating unilateral small branchial arches (BA; red arrowhead). B: Tadpole with bilateral reduction in BA and jaw (red arrowheads). C: Tadpole with bilateral reduction in BA and jaw (red arrowheads), and eyes with thickened pigmented optic nerves (blue arrowhead) that are joined across the midline. The left otolith is missing (orange arrowhead). D: Tadpole with abnormal right BA and jaw (red arrowhead), an eye with thickened pigmented optic nerve (blue arrowhead), and a missing right otolith (orange arrowhead). E: Tadpole with pigmented optic nerves (blue arrowheads). F: Tadpole with missing left olfactory pit (yellow arrowhead), left eye fused to the brain (blue arrowhead), and malformed left otocyst (orange arrowhead). G: Tadpole with its right olfactory pit fused to the brain (yellow arrowhead), and a malformed right eye (blue arrowhead). H: Tadpole with a missing right otolith (red circle); normal left otolith circled in green. I: Profile of tadpole lacking jaws (red arrowhead). Inset: normal profile. Image published in: Vandenberg LN et al. (2011) Copyright © 2011. Image reproduced with permission of the Publisher, John Wiley & Sons. Experiment + Assay Source Phenotypes and Disease Xla Wt + atp6v0c + NF45 (whole-mount microscopy) Fig S3 A Anatomical Phenotype abnormal branchial arch skeleton morphology decreased size of the branchial arch skeleton Xla Wt + atp6v0c + NF45-48 (whole-mount microscopy) Fig S3 B Anatomical Phenotype abnormal branchial arch skeleton morphology decreased size of the branchial arch skeleton decreased size of the Meckel's cartilage Xla Wt + atp6v0c + NF45-48 (whole-mount microscopy) Fig S3 C Anatomical Phenotype absent inner ear decreased size of the branchial arch skeleton decreased size of the Meckel's cartilage fused optic stalks increased pigmentation in the optic stalk increased thickness of the optic stalk Xla Wt + atp6v0c + NF45-48 (whole-mount microscopy) Fig S3 D Anatomical Phenotype absent inner ear decreased size of the branchial arch skeleton decreased size of the Meckel's cartilage increased thickness of the optic stalk Xla Wt + atp6v0c + NF45-48 (whole-mount microscopy) Fig S3 E Anatomical Phenotype increased pigmentation in the optic stalk Xla Wt + atp6v0c + NF45-48 (whole-mount microscopy) Fig S3 F Anatomical Phenotype abnormal inner ear morphology absent olfactory pit fused eyes Xla Wt + atp6v0c + NF45-48 (whole-mount microscopy) Fig S3 G Anatomical Phenotype abnormal eye morphology abnormal olfactory pit morphology Xla Wt + atp6v0c + NF45-48 (whole-mount microscopy) Fig S3 H Anatomical Phenotype abnormal inner ear morphology Xla Wt + atp6v0c + NF45-48 (whole-mount microscopy) Fig S3 I Anatomical Phenotype absent Meckel's cartilage Larger Image Printer Friendly View

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