Fig 1.
Pedigree drawing of family PKCC074.
The haplotypes of five adjacent chromosome 16q markers are shown with alleles forming the risk haplotype shaded black and alleles not co-segregating with cataract are shown in white. Squares: males; circles: females; filled symbols: affected individuals; the double line between individuals: consanguineous matting; and a diagonal line through a symbol: deceased individual.
Fig 2.
Slit-lamp photograph of individual 12 of family PKCC074.
The photograph illustrates nuclear cataracts in the affected individual.
Table 1.
Two-point LOD scores of chromosome 16q markers for family PKCC074.
The asterisk indicates the marker included in the genome-wide scan.
Fig 3.
Sequence chromatograms of HSF4 in family PKCC074.
A) Unaffected individual 7 homozygous for wild-type allele; B) unaffected individual 8 heterozygous and C) affected individual 9, homozygous for c.433G>C variation that results in a p.Ala415Pro. D) Sequence alignment of amino acids illustrating the conservation of the Ala145 and its neighboring amino acids. The organisms in green and purple are primates and placental mammals respectively.
Fig 4.
Expression pattern of Hsf4 in mouse lens at different developmental time points.
The expression levels were normalized with Gapdh and the expression of Hsf4 at embryonic day 15 (E15) was chosen as a reference point to compare the relative expression at different developmental time points. The x-axis and y-axis represent developmental time points and relative expression of Hsf4 mRNA, respectively.
Fig 5.
Sub-cellular localization of wild-type and mutant HSF4b (p.Ala145Pro) in HeLa cells.
The cells were transfected with wild-type (A, C) and mutant HSF4b (B, D) constructs fused with C-terminus FLAG. The monoclonal anti-FLAG antibody was used to detect the FLAG-tagged wild-type and mutant HSF4b proteins. Nuclei were stained with DAPI. The wild-type (panels A and C) and mutant HSF4b (panels B and D) illustrate a nuclear localization pattern. The panels (A and B) and (C and D) represent 20× and 100× magnifications, respectively.
Fig 6.
A schematic representation of the protein structure and causal variants identified in HSF4.
Mutations previously identified in HSF4 responsible for autosomal dominant cataracts are shown in brown, mutations responsible for autosomal recessive cataracts are shown in blue. The p.Ala145Pro mutation shown in red was identified in the study. Note: In addition to variants presented in the figure, a splice site variant (IVS5 c.233-1G>A) in HSF4 has been reported for autosomal dominant cataracts by Cao and colleagues [38].