Fig 3. PCR validation of Alu insertion identified by in silico analysis in patients from the discovery cohort.

A) PCR amplification using primers spanning exon 9 (top) and nested PCR using Alu-specific primer (bottom). The 1,194 bp amplicon containing the Alu insertion (arrow) is present strongly in the homozygous sample and weakly in the heterozygous samples (top); the Alu insertion-specific amplification (491 bp, bottom) confirms the presence of the Alu insertion. B) Pedigree of patient D379_148, carrying a missense mutation (p.Gly16Arg) and the Alu insertion mutation. C) Evolutionary conservation of glycine 16, mutated in the patient D379_148. D) Protein domains in MAK and location of the p.Gly16Arg change. The mutation annotations are based on the NM_001242957 transcript, where A from the ATG start codon is designated as a +1 position.