Figure 3.

Molecular analysis. (A) Gel electrophoresis of the RT-PCR products from HEK293T cells transfected with minigene plasmids carrying exon 5 to exon 6 of LTV1 and either the WT allele or the c.503A > G mutation. The mutant form of the plasmid has two transcripts, corresponding to the correctly spliced isoform (1) and an aberrantly spliced isoform (2). This non-canonical transcript (2,3) is present only in cells transfected with minigenes bearing the mutation, as specifically shown by the use of an isoform-specific primer (CR 7326). (B) Sanger sequencing of the two transcripts, from the WT minigene (top) and the one carrying c.503A > G (bottom). (C) Schematic view of the LTV1 minigene, depicting the position of the variant identified and the splicing event resulting from its presence, as well as the position of the PCR primers used.