Fig 1.
Pedigrees of eight ET families that were whole genome sequenced.
Pedigrees for families (A-H) that were whole genome sequenced are shown. The generation in each pedigree is shown by roman numerals. The proband is indicated by an arrowhead. A ‘*’ symbol indicates subjects that were whole genome sequenced. Below each subject with DNA avaliable for genetic analysis the subject ID is indicated. Symbol shading is as follows: definite ET, symbols completely black; probable ET, symbols half vertical black fill; possible ET, symbols with a quadrant in black; and unaffected clear symbol. To protect the identity of participants in families the gender and birth order were changed in order to disguise their identities.
Table 1.
Clinical characteristics of affected ET individuals and unaffected family members that were whole genome sequenced in eight families.
Fig 2.
Analysis workflow for analysis using MM-KBAC.
The analysis workflow for WGS data is shown with population database filtering, analysis methods and annotation.
Table 2.
Variants identified in families co-segregating with ET based on MM-KBAC analysis of rare variants by variant type.
Table 3.
Synonymous variants in enhancer and splicing regions identified in families co-segregating with ET based on MM-KBAC analysis of rare variants.
Table 4.
Variants located within TFBS identified in families co-segregating with ET.
Table 5.
Rare CNVs segregating with ET in families.
Fig 3.
Behavioral manifestations of nervous system dysfunction in a slit Drosophila model.
(a) Climbing response during lifespan. The climbing assay was assessed as the time taken for the first fly to climb 10.0cm. The mean climbing index + SEM as a function of age is shown for each independent mutant slit line and the wildtype line. Each point represents the mean of 10 flies. Flies expressing the mutant slit (p.Val1187Leu) compared to wildtype slit displayed significantly slower climbing (p<0.05) throughout lifespan. (b) Survival assays in slit lines. A total of 100 virgin flies per line were sex segregated within 4h of eclosion and maintained in small laboratory vials (n = 20 per vial) containing fresh food in a low-temperature incubator at 25°C and 40% humidity on a 12/12h dark/light cycle. The flies were transferred to fresh food vials every 3–4 days and mortality recorded. Significant differences in lifespan were detected between flies expressing the mutant slit (p.Val1187Leu) compared to wildtype slit (p<0.0001).
Fig 4.
Cav3.1 electrophysiology for mutant and wildtype channels at room temperature and at near physiological temperature.
(a) Current-Voltage relationship of wildtype and mutant Cav3.1 channels. (b) Time to peak with wild type and mutant Cav3.1 channels. (c) I-V relationship of wild type and mutant Cav3.1 channels. (d) Steady state inactivation of wild type and mutant channels.