Figure 2

Deep intronic mutation within GPR143 intron 5.

(a) WES coverage data around GPR143 exon 6 of the SNV calls of interest of the family members shown using the Integrated Genomics Viewer. The arrowhead denotes mutated bases and the arrow indicates the direction of transcription.(b) Sanger sequence analysis of GPR143 intron 5 and exon 6. Arrowheads denote the mutated base. Electropherogram of the affected siblings (II:1 and II:2) showing a hemizygous c.659-131 T > G GPR143 mutation. Electropherogram of their mother’s (I:2) sequence showing the heterozygous c.659-131 T > G GPR143 mutation that was undetectable in their father’s (I:1) DNA.(c) Predictions of splice sites in the wild-type and mutated GPR143 genomic sequences. Diagram of the GPR143 region comprising exons 5 and 6. Closed arrow indicates intron 5 (IVS5). Arrowheads indicate the mutated base. Gray boxed sequences represent the 41-bp pseudoexon created by the mutation. The scores calculated using MaxEntScan, NNSPLICE and Human Splicing Finder are displayed below each splice site. A higher score predicts a strong splice site. The values provided in parentheses indicate the score ranges in each algorism. Note that scores of a potential cryptic splice donor site created by the mutation was almost the same levels as those of authentic splice donor and acceptor sites for IVS5.