Figure 4. Versican (VCAN) transcript analysis. A: Ethidium bromide-stained agarose gel of reverse transcription- polymerase chain reaction (RT–PCR) products obtained with primer pairs 7F/8R for amplification of the versican V0 mRNA from cultured lymphoblasts of one affected individual (V.5) and one control unaffected subject (V.4). The results indicate that both a wild-type (band of 372 bp) and a 39-nt deleted (band of 333 bp) cDNA fragment of the V0 transcript are present in the affected patient harboring the intron 7 splice acceptor mutation of the VCAN gene (c.4004–2A>T). The nucleotide change at the 3′ acceptor splice site of intron 7 of the VCAN gene was analyzed using the NNSplice program for prediction in alteration of splicing junctions, and the splice-site scores (SSSs) for a normal site (SSS-nor), a mutated site (SSS-mut), and a cryptic site (SSS-crypt) were calculated [28]. B: Partial sequence chromatogram of this aberrant V0 transcript lacking the first 39 bp of VCAN exon 8.