Fig. 2

Expression analysis in transgenic zebrafish. A) Example of a zebrafish with a GFP positive heart. B) Copy number analysis of integrated human PTDSS1, GFP and mCherry by q-PCR on gDNA isolated from individual GFP- fluorescing zebrafish. C) Comparison of DNA copy number (top panel) and RTq-PCR expression analysis (bottom panel) for individual zebrafish where DNA and RNA were extracted from the same samples. D) Comparison of endogenous zebrafish ptdss1 with human PTDSS1 showing significantly higher levels of endogenous transcripts over WT and mutant transgene transcripts (p=0.01 and 0.008 respectively).

Fig. 3

Analysis of transgene expression in transgenic zebrafish. A) Immunoblotting showing mCherry and ERK (control) in extracts from 3 d.p.f. transgenic zebrafish (either wild-type, Wt, or Q353R mutant, Mut) and non-transgenic (NT) zebrafish as well as HeLa cells transfected with the same beta-actin (BA) constructs. B) Similar western blot including transgenic zebrafish at 7 d.p.f and 14 d.p.f.

Fig. 4

Imaging of zebrafish skeletons using alizarin red staining. Spinal cord, ribs and fin rays in A) a non-transgenic zebrafish; B) a transgenic zebrafish showing spine/tail kink; C) a transgenic zebrafish with scoliosis. E) Graphical representation of the distribution of skeletal defects seen for each transgenic line tested.

Acknowledgments

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