Fig. 3.

Minigene splicing assay for the variant c.706-2A>C. a The pSLP3 vector contains a functional intron and a multiple cloning site within the two exons SD and SA. PCR fragments containing the CYP26C1 genomic sequence intron3-exon4-intron4 wt or c.706-2A>C were cloned within the multiple cloning site. The expected splicing events are indicated in the scheme. b Agarose gel electrophoresis of the RT-PCR amplicons. Forward SD6 and reverse SA2 primers were used for RT-PCR of cDNA generated from U2OS cells transfected with pSPL3 empty vector, pSPL3-CYP26C1 intron3-exon3-intron4 wt, or pSPL3-CYP26C1 intron3-exon4-intron4 c.706-2A>C. MCS multiple cloning site