FIGURE 1

Generation of hGNE^M743T/WT transgenic lines. (A) A schematic representation of the p(-1.9gne:hGNE^M743T/WT; insulin:EGFP) plasmid. Genes, promoters, LTRs, polyadenylation signal and restriction sites are indicated. (B) The hGNE transgenic lines develop normally. The figure presents brightfield images of a WT (top), hGNE^WT transgenic (middle), and hGNE^M743T transgenic (bottom) larvae at 8 dpf. Scale bar = 1 mm. (C) RT-PCR analysis of hGNE mRNA expression in hGNE^M743T and hGNE^WT transgenic lines. Agarose gel electrophoresis of RT-PCR amplification products of the hGNE gene (top) and eef1a1a (bottom) as the internal control. Both the hGNE^M743T (Mut) and hGNE^WT (WT) cDNA samples show the expected amplification in all tested time points and tissues. Non-transgenic WT zebrafish cDNA (WT ZF cDNA) and genomic DNA from transgenic fish (Tg DNA) served as controls for each target. (NTC) no template control, (M) Muscle, (Br) Brain tissue.