Fig. 4

a RT-PCR products of the c.3286 + 5G > A in pSPL3 minigene constructs. Lane 1: the 1000 bp marker. Lane 2 and lane 3: the splicing aberrant band. Lane 4 and lane 5: the normal band. Lane 6 and lane 7: empty vector. Lane 8: blank control. b Splicing schematic representation of the mini gene vectors used for the in vitro splicing assay. The wild type transcripts have three diverse bands, band 1(407-bp), band 2(288-bp) and band 3(263-bp). Band 1 was the products of exon 24 and exon 25. Band 2 have exon 25. There are no exon in the band 3, which was only the sequence of the pSPL3 vectors. The band 1 was absent in mutation type .The mutation c.3286 + 5G > A led to aberrant splicing that exon 24 was skipping in the mini gene splicing assay. c Direct sequencing after gel extraction of the band 1, band 2 and band 3