PMC9982009_01
Preliminary experiments. (A) Sanger sequencing results for validation of the DUOX2 mutations in c.1588A>T (p.K530X) and c.2654G>T (p.R885L) in family members. The black background indicates the targeted mutation sites. Arrows denote mutations. (B) Pedigree haplotyping analysis based on informative SNPs flanking DUOX2. The arrow indicates the proband. Circles and squares indicate females and males, respectively. The filled symbol represents the affected patient. Half-filled symbols represent DUOX2 mutation carriers. Haplotypes are represented by colorful bars combined or not combined with diagonal lines. The dark green bar indicates a paternal wild-type haplotype. The light green bar combined with diagonal lines denotes a paternal mutation: DUOX2, c.2654G>T (p.R885L). The dark orange bar denotes a maternal wild-type haplotype. The light orange bar combined with diagonal lines indicates a maternal mutation: DUOX2, c.1588A>T (p.K530X). (C) Pedigree haplotypes around chromosomal breakpoint regions. The arrow indicates the proband. Circles and squares indicate females and males, respectively. Half-filled symbols represent derivative chromosome carriers. In ChrX, the dark green bar indicates the normal paternal ChrX haplotype. The dark orange bar denotes the normal maternal ChrX haplotype. Light orange bars with diagonal lines represent the derivative ChrX haplotype. In Chr18, the dark and light green bars indicate a pair of paternal normal Chr18 haplotypes in two homologous chromosomes. The dark orange bar represents the normal maternal Chr18 haplotype. The light orange bar combined with diagonal lines denotes the derivative Chr18 haplotype.
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F1
After thyroxine treatment, his TSH levels decreased to 2.22 mIU/L (11 April 2017) (Supplementary Figure S1).
Linkage analysis and mutation detection in embryos. (A) Haplotypes around DUOX2 of the family members and embryos. Diamonds indicate embryos, and the interpretation of other symbols and bars is explained in the legend of Figure 1B. E1 inherited both wild-type haplotypes from parents. E2 inherited the paternal mutation and maternal wild-type haplotype. E3 inherited the paternal wild-type haplotype and maternal mutation. (B) The mutations in embryos were identified by Sanger sequencing. The results were consistent with those of the SNP-based linkage analysis.
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F2
Karyotype analysis of the proband and his parents revealed normal results (Supplementary Figure S2).
CNV tests and SNP-based linkage analysis for RecT detection in embryos. (A) Embryonic CNV results. E1 was unbalanced. E2 and E3 were chromosomally balanced. (B) Pedigree haplotyping around chromosomal breakpoints. The interpretation of symbols and bars is explained in the legend of Figure 1C. E2 inherited its two wild-type X chromosomes and two wild-type 18 chromosomes from their parents. It was female and free of RecT. E3 inherited a derivative ChrX and derivative Chr18 from the mother. It carried RecT between ChrX and Chr18.
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F3
The gene variant pedigree of the family was constructed (Supplementary Figure S3).
Prenatal diagnostic results. (A) Sanger sequencing results for fetal DUOX2 mutations. Red boxes represent targeted mutation sites. The arrow indicates the DUOX2, c.2654G>T (p.R885L) mutation. (B) Amniotic fluid karyotyping of the fetus, revealing a normal karyotype. (C, D) Results of SNP array-based comprehensive chromosome screening.
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F4
The results of CNV sequencing (0.6x sequencing depth) revealed that the proband exhibited an Xq28-q28 duplication (5.59 Mb x 2) and 18q22.2-q23 deletion (9.85 Mb x 1) and the CNVs of the parents were normal (Supplementary Figure S4).