XB-IMG-79038
Xenbase Image ID: 79038
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Supplemental Figure 1. The C-terminal tail of fus is dispensable for splicing. (A) fus genomic locus showing the full gene model in the top panel, and a close up of the last four exons in the bottom. The C-terminal region contains eight conserved amino acid residues (orange) that have been linked to fALS in humans and seven of these are encoded in the last exon 15. The splice blocking MO fus-fALS-MO5 (red) targets the donor splice site in exon 14. (B) Single embryo RT-PCR on two control embryos or fus-fALS morphants at stage 25. (B, Top panel) Missplicing of the last splice junction of fus resulting either in intron inclusion (fus-fALS-C), causing premature stop or use of two cryptic splice donor sites in exon 14 (fus-fALS-B and fus-fALS-A) that cause a frame shift. In all cases the result is C-terminal truncated forms of fus without the C-terminal region containing the residues mutated in fALS mutations. (B, lower panels) normal splicing and expression of fgfr2 and fgf8 in fus-fALS morphants. Internal control was odc. (C) fus-fALS morphants develop normally at least until stage 41. (D) fus-fALS morphants are immobile after hatching (stage 25, left barplot). As development proceeds fus-fALS morphants gain mobility, but are still impaired compared to normal embryos (stage34-38, right barplot). Normal swimming was scored as ability to swim for at least one second after being prodded. Swimming for shorter duration was scored as impaired. Image published in: Dichmann DS and Harland RM (2012) Copyright © 2012. Image reproduced on with permission of the Publisher, Cold Spring Harbor Laboratory Press. This is an Open Access article.
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