<i>In vitro</i> functional analysis of mutation c.1242+1G>A and exon 11 splicing analysis in normal fibroblasts.

<p>Panel A: RT-PCR analysis of the <i>HEXB</i> mRNA in cells transfected with normal (pcDNA3HEXBN) and a minigene containing mutation c.1242+1G>A (pcDNA3HEX1242) MW: 1 kb Plus DNA Ladder. Panel B: Schematic representation of the effect of the novel mutations on the splicing process. Sequencing analysis of RT-PCR products showed that the presence of c.1242+1G>A mutation determines the skipping of 73 nt of exon 10. In addition the presence of a 3′ cryptic splice site (present in both normal and mutant minigenes) determines the skipping of 112 nt in exon 11(denoted as red box) in cells transfected with both normal and mutant minigenes. Panel C: RT-PCR analysis of the <i>HEXB</i> mRNA in normal fibroblasts. MW: 1 kb Plus DNA Ladder.