Fig. 1

Characterisation of vhl−/− zebrafish. A) Gross morphology of 4 and 5 dpf vhl larvae highlighting the absence of a swim bladder, plus yolk and pericardial oedema in vhl-/- larvae. Scale bar = 2 mm. B) Phenotype-Genotype correlations in offspring generated from vhl ± incrosses. DNA sequencing of PCR amplicons from genomic DNA confirmed a positive correlation between swim bladder uninflation and vhl−/− genotype. This strategy was used to select vhl−/− larvae for all further experimentation. C) Sequencing chromatograms illustrating the single nucleotide polymorphism (C > T) in the vhl sequence resulting in a premature STOP codon. D) Real time PCR analysis of vhl expression in vhl−/− larval eyes in comparison to siblings (+/− or +/+). ns = not significant when p < 0.05. E) Reverse transcriptase PCR highlighting vhl expression in vhl−/− and sibling eyes at 5 dpf.