Figure and table captions
Figure1 Pedigree with Axenfeld-Rieger syndrome in the proband. Family members and disease-causing mutations are presented. Normal individuals are shown as clear circles (females) or squares (males), whereas the affected individual is shown as a filled square with an arrow indicating the proband (III:2) with the missense variant of the FOXC1 gene: c.246C>A. “M” indicates the mutant allele of FOXC1 (mutant type), whereas “+” indicates the normal allele (wild type).
Figure 2 Representative anterior segment photography of patient III:2 from both eyes. A and B represent the right eye and left eye, respectively. C shows systemic and ocular characteristics of patient III:2. D shows developmental malformation of the teeth, while E shows the outward projection of the navel.
Figure 3 Pyrogram profiles for mutation verification by dideoxy DNA sequencing shows overlapping peaks at nucleotide position 246 (red arrow) due to the missense variant of the FOXC1 gene: c.246C>A. in the proband (III:2), while the variant was not detected in the proband’s parents (II:1, II:2), his brother (III:1) or his grandmother (I:2). Unfortunately, DNA samples of his grandfather (I: 1) were not collected.
Figure 4 Multiple sequence alignment of the region of the FOXC1 protein surrounding the novel p.S82R mutation in various species. The serine(s) residue (indicated with a red strip-type frame) was highly conserved among all species analyzed.
Figure 5 The results showed that the 82nd amino acid of the wild type was serine, which changed to arginine after mutation, leading to an uncharged amino acid (A, wild type) change to a positively charged amino acid (B, p.S82R).
Table 1 Clinical features of the pedigree members. Abbreviations: VSD, ventricular septal defect
Table 2 Characteristics of the FOXC1 variant in the proband and analysis of its disease-causing effects. Abbreviations: c, variation at the cDNA level; p, variation at the protein level; S82R, serine substitution conserved arginine at codon 82; hetero, heterozygote; AD, autosomal dominant inheritance.
Table 3 PCR primers and PCR product size