Figure 3. Pyrosequencing assay. (A) Schematic representation of the SMC1A protein showing relative positions of the mutations on the functional domains. Boxes indicate the N- and C-terminal NTPase-binding cassettes (ABC) (amino acids 4–148 and 1117–1220, respectively) and the crucial hinge motif (amino acids 513–628), while the black line represents the coiled-coil domains. The position of the coding SNP rs1264011 at the N-terminus of the protein is indicated. Mutation-specific pyrosequencing assays were performed for each of the six investigated CdLS patients, and a specific assay for the coding SNP rs1264011 was used for the 15 heterozygous controls. (B) Pyrosequencing results. Expression levels of the wild type (allele 1) and mutant (allele 2) SMC1A alleles in six patients (Pt). In four patients, RNA expression analysis of both peripheral blood (PB) and LCLs are shown. (C) Histograms of pyrosequencing analysis data. Patients’ (Pt) wild type allele expression is shown in light gray and mutant allele expression is in dark gray. For comparison, the relative allele expression in controls (1:1) is indicated with a horizontal black line.