Table 2

DHH gene variant functional activity and second curation

Patient ID	DHH variants		Curation before functional	% of WT activity (mGli1)	Adjusted p value (mGli1)	% of WT activity (mPtch1)	Adjusted p value (mPtch1)	Final curation
1	NM_021044.2:c.491G>C:p.Arg164Pro	Homozygous	LP	1%	<0.0001	4%	0.0398	LP
2	NM_021044.2:c.680C>T:p.Ala227Val	Compound	VUS-3a	−1%	<0.0001	−20%	0.0059	LP
	NM_021044.2:c.782C>T:p.Pro261Leu	heterozygote	VUS-3a	2%	<0.0001	4%	0.0377	LP
				Combined 2 %	<0.0001	Combined 11 %	0.409
3	NM_021044.2:c.746G>A:p.Arg249His	Compound	VUS-3a	26%	0.0002	13%	0.0448	LP VUS-3a
	NM_021044.2:c.508G>A:p.Gly170Ser	heterozygote	VUS-3a	47%	0.0096	31%	0.2342	LP VUS-3a
				Combined 46 %	0.0091	Combined 41 %	0.409
4	NM_021044.2:c.508G>A:p.Gly170Ser	Heterozygous	VUS-3a	47%	0.0096	31%	0.2343	VUS-3a
				Combined 90 %	0.9956	Combined 65 %	0.9155	VUS-3a
5	NM_021044.2:c.583C>T:p.Arg195Trp	Heterozygous	VUS-3a	16%	<0.0001	2%	0.0334	VUS-3a
				Combined 98 %	0.9999	Combined 72 %	0.9777	VUS-3a
6	NM_021044.2:c.80G>A:p.Arg27Gln	Heterozygous	VUS-3a	67%	0.9956	78%	0.9957	VUS-3c (potentially benign)
				Combined 65 %	0.1674	Combined 53 %	0.6787	VUS-3c (potentially benign)

Results from co-culture functional assay are shown, including percentage of WT activity and p values for both mGli1 and mPtch1 read-outs. Where variants were found as compound heterozygous or heterozygous, functional read-outs for combined alleles are also shown. Curation outcomes for each variant taking these functional outcomes into account are also shown.
DHH, desert hedgehog; LP, likely pathogenic; VUS, variant of uncertain significance; WT, wild type.