Fig 2: Missense Variant c.1864A > G (p.Met622Val) in ANO10 Causes 2 Alternative Splicing Transcripts(A) Gel electrophoresis of fibroblast cDNA products spanning exon 12 of ANO10 shows 2 abnormal splicing events in patient fibroblasts (P3) relative to wild-type control fibroblasts (C1 and C2). (B) Sanger sequencing chromatographs showing sequence coverage of wild-type ANO10 cDNA and 2 transcript variants. Transcript variant 1 shows the use of wild-type splicing enhancers, which incorporates exon 12 containing c.1864A > G (p.Met622Val). Transcript variant 2 results in skipping of exon 12. No wild-type sequence could be detected in patients.

Fig 3: Collation of ANO10 Variants Found in the LiteratureShowing the percentage (number of individuals) of cases reported with the indicated common clinical symptoms of SCAR10 for each variant and/or variant combinations as indicated. 100 = 100% of the cases that reported the phenotype were positive; 0 = 0% of the cases that reported the indicated phenotype were positive; NA = not reported in any individual within the group.

Fig 4: Intronic Variant c.1163-9A > G in ANO10 Causes 2 Alternative Splicing Transcripts(A) Gel electrophoresis of fibroblast cDNA products spanning exon 7 of ANO10 shows 2 abnormal splicing events in patient fibroblasts (P1) and blood (P2) relative to wild-type control fibroblasts (C1 and C2) or control blood (C3 and C4). (B) Sanger sequencing chromatographs showing sequence coverage of wild-type ANO10 cDNA and 2 transcript variants. Transcript variant 1 shows the use of a cryptic splice site acceptor that incorporates 8 base pairs of the adjacent intron 6. Transcript variant 2 results in skipping of exon 7. No wild-type sequence could be detected in patients. (C) Schematic representation of the abnormal splicing events in ANO10 generated by the homozygous intronic variant c.1163A>G–9. Wild type shows a normal relationship between splice site donor and acceptor sites, while the variant introduces a cryptic splice site acceptor, which is preferred over wild-type splice site acceptor (transcript variant 1) and retains 8 base pairs of intron 6, or is ignored altogether (transcript variant 2) leading to skipping of exon 7.