Fig. 5

Variants that were not detected by lrWGS. A Pedigree of family F6440 with three stillbirths all presenting with profound global developmental delay, microcephaly, epilepsy, and scoliosis. B Clinical image of an affected individual (III:10) highlighting microcephaly. C EasyLinkage analysis showing a single locus at chr9:31537680–79306780 with LOD of 4. D Screenshot of Bionano analysis output identifying an insertion (chr9:33,266,774–33,271,717) disrupting CHMP5. E RT-PCR experiment showing 91 bp insertion in intron 2 (I2). F Western blot experiment showing reduction of CHMP5 protein levels in two affected individuals compared to two independent controls. Alpha-Tubulin was used as loading control. G Relative quantification of CHMP5 protein levels showing ~ 80% reduction compared to controls. H Pedigree of family F8280 with two siblings affected with SHFM. I Clinical image of limb malformation of affected individual (IV:1). J Chromosomal microarray output from family F8280 showing heterozygous duplication in the known SHFM locus that was absent in the parents