Fig. 2

In vitro splicing assay using hybrid minigene construct. a RT-PCR amplified products of hybrid minigene transcripts. As shown in the gel file, a comparatively small band was present in the wild type construct both in HEK293T and HeLa cells. In contrast, in the minigene derived from the patient sequence, only a relatively large band was detected in both cell lines, which corresponds to a 104 bp insertion between exon 11 and exon12. b The RT-PCR product containing the 104 bp insertion between exon 11 and exon 12 was obtained from the minigene encoding the intronic mutation (b top). On the other hand, the product containing exon 11 and exon 12 could be obtained from the hybrid minigene encoding the wild type genomic DNA (b bottom)