Fig. 4

Schematic showing the effect of the splicing mutation. a. The retrotranscription of RNA extracted from HEK293 cells transfected with WT/Mutated minigene vectors (see the methods) leads to amplicons of different size: a full chimeric amplicon (665 bp) and a shorter chimeric cDNA fragment (434), both sequenced in c; b. cDNA amplification with an internal primer designed on the exon 5, shows that both the WT and the mutated RNA produce a normal amplicon (here not quantified): this is partially confirmed by the “faint” band in the lane of mutated cDNA; d: diagram showing the exon skipping mechanism induced by the presence of the IVS5 + 1G > A mutation