Fig. 1: DNA sequencing of ten unrelated MAM-negative individuals revealed inactivating mutations in EMP3.

a Exome sequence alignments over the EMP3 gene region demonstrate four inactivating mutations in five MAM-negative individuals. EMP3 was the only candidate gene that passed our filtering strategy with predicted loss of function mutations found in all tested MAM-negative samples. Upper panel (chr19; 48,822,471 to 48,837,471) shows a complete deletion of EMP3 in P9 (blue box) revealed by a lack of coverage over any targeted exons as compared to control, although flanking genes CCDC114 and TMEM143 show sequencing coverage. Lower panel (chr19; 48,828,000 to 48,834,500) shows homozygous nonsense mutation (c.123C > G; p.Tyr41Ter) in P2 (brown line); deletion of EMP3 exon 4 in P8 (pink box) and P5 (data not shown); deletion of EMP3 exon 5 in P4 (orange box). b Sanger sequencing was used to confirm these mutations and identify EMP3 mutations in a further five MAM-negative individuals and deletion breakpoints were identified where appropriate (summarised in Supplementary Table 5). EMP3 inactivating mutations in all patients (P1–P10) are depicted on the gene schematic, with dark blue areas depicting coding exons, light blue areas UTRs and deleted regions depicted by dashed red lines.