<p>(A) CLUSTAL alignment indicates that zebrafish Cln3 amino acid sequence (second row) is 62% similar and 49% identical to human CLN3 (first row). Black background indicates identical residues shared between human and zebrafish, grey background indicates similarity, ‘-’ indicates a gap in the alignment. (B) RT-PCR using primer pairs AF1-AR1 and AF1-AR2 (see C), followed by sequencing, confirmed that the <i>cln3</i> SPL MO affected mRNA splicing. Lanes one and two show the PCR product from cDNA made from <i>cln3</i> SPL MO injected embryos and WT embryos respectively. Arrow in lane three points to the positive control, which is the PCR product generated using plasmid containing <i>cln3</i> mRNA as a template. (C) i) the gene structure of WT <i>cln3</i> showing the relative position of the PCR primers used. ii) The WT mature mRNA. iii) Splice variants caused by injection of <i>cln3</i> SPL MO. Key: lines, introns; boxes, exons; white boxes, untranslated regions; red asterisk, stop codon.</p
