Fig. 3

In vitro and in vivo functional analysis of IFT140 c.634G>A; p.Gly212Arg. a Schematic of the human IFT140 locus at chr16:1,510,427-1,612,110 (hg19; top) and translated protein (bottom). Exons are depicted as green boxes; untranslated regions are shown as white boxes (NM_014714.3). Protein schematic (blue; NP_055529.2) indicates predicted WD40 repeat (WD40) and tetratricopeptide-like helical domains (TPR). IFT140 c.634G > A; p.Gly212Arg location is indicated with a red star. b RT-PCR from proband lymphocyte cell lines indicating aberrant mRNA splicing. IFT140 cDNA was amplified with primers flanking the exon 6 canonical splice donor site (black arrows). Sequence chromatograms from purified PCR product indicates that the majority of IFT140 message is missing exon 6 (bottom). c Lateral (top) and dorsal (bottom) views of ift140 morphants exhibiting gastrulation defects. d In vivo complementation studies. Live embryo batches were scored to assay variant pathogenicity. Class I, modest shortening of the body axis and reduction in size of anterior structures; class II, severe shortening of the body axis and decreased anterior structures accompanied by broadening and/or kinking of the notochord and thinning of the somites. To compare pairs of embryo batches, χ ² tests were used; p < 0.0001 indicated with (asterisk)