Genetic analysis of pedigree.

<p>a) Pedigree examined in this study. Only individuals from generations III and IV were available for genetic analysis. b) Location of the mutation NM_020919.3:c.3512+1G>A at the boundary of the 21st intron of the <i>ALS2</i> gene and c) confirmation by Sanger sequencing. d) RT-PCR of total RNA isolated from patient (IV-4 labeled as ALS2) and two control fibroblast cells (labeled BJ and CV), visualized on a gel alongside an Invitrogen 1 kb+ ladder. Three splicing transcripts corresponding to the three bands (102 bp encoding p.Ser1116_Thr1170del [red], 200 bp encoding p.Pro1148fs [blue] and 267 bp encoding the normal protein [black]) were confirmed by Sanger sequencing and illustrated in the figure.</p

Top four most significant genes associated with endometrial cancer risk in the Swedish population.

+<p>P value is based on 5000 permutations in AML test for the specific gene.</p

Sub-complex AML test for genetic association between the tag SNPs in the NCOA2 complex and endometrial cancer risk in the validation analysis.

<p>Sub-complex AML test for genetic association between the tag SNPs in the NCOA2 complex and endometrial cancer risk in the validation analysis.</p

Global P values of AML tests for genetic association between the tag SNPs in the ER cofactor complexes and endometrial cancer risk in the Swedish population.

<p>P-values were based on 5000 permutations.</p

Sub-complex AML tests for genetic association between the tag SNPs in the histone acetylation & methylation and endometrial cancer risk in the Swedish population.

<p>Sub-complex AML tests for genetic association between the tag SNPs in the histone acetylation & methylation and endometrial cancer risk in the Swedish population.</p

QQ plot for trend tests of 685 ER cofactor SNPs associated with endometrial cancer risk in the Swedish population.

<p>QQ plot for trend tests of 685 ER cofactor SNPs associated with endometrial cancer risk in the Swedish population.</p

Endometrial cancer sample sets and sources included in initial and validation analysis.

<p>Endometrial cancer sample sets and sources included in initial and validation analysis.</p

List of top five SNPs in the combined analysis in the NCOA2 and CREBBP gene respectively.

+<p>Real genotyped SNPs.</p

Genome-Wide Linkage, Exome Sequencing and Functional Analyses Identify <i>ABCB6</i> as the Pathogenic Gene of Dyschromatosis Universalis Hereditaria

<div><p>Background</p><p>As a genetic disorder of abnormal pigmentation, the molecular basis of dyschromatosis universalis hereditaria (DUH) had remained unclear until recently when ABCB6 was reported as a causative gene of DUH.</p><p>Methodology</p><p>We performed genome-wide linkage scan using Illumina Human 660W-Quad BeadChip and exome sequencing analyses using Agilent SureSelect Human All Exon Kits in a multiplex Chinese DUH family to identify the pathogenic mutations and verified the candidate mutations using Sanger sequencing. Quantitative RT-PCR and Immunohistochemistry was performed to verify the expression of the pathogenic gene, Zebrafish was also used to confirm the functional role of ABCB6 in melanocytes and pigmentation.</p><p>Results</p><p>Genome-wide linkage (assuming autosomal dominant inheritance mode) and exome sequencing analyses identified <i>ABCB6</i> as the disease candidate gene by discovering a coding mutation (c.1358C>T; p.Ala453Val) that co-segregates with the disease phenotype. Further mutation analysis of <i>ABCB6</i> in four other DUH families and two sporadic cases by Sanger sequencing confirmed the mutation (c.1358C>T; p.Ala453Val) and discovered a second, co-segregating coding mutation (c.964A>C; p.Ser322Lys) in one of the four families. Both mutations were heterozygous in DUH patients and not present in the 1000 Genome Project and dbSNP database as well as 1,516 unrelated Chinese healthy controls. Expression analysis in human skin and mutagenesis interrogation in zebrafish confirmed the functional role of <i>ABCB6</i> in melanocytes and pigmentation. Given the involvement of <i>ABCB6</i> mutations in coloboma, we performed ophthalmological examination of the DUH carriers of <i>ABCB6</i> mutations and found ocular abnormalities in them.</p><p>Conclusion</p><p>Our study has advanced our understanding of DUH pathogenesis and revealed the shared pathological mechanism between pigmentary DUH and ocular coloboma.</p></div

Number of mature melanocytes in the head region in the morphants and rescued embryos (5 dpf).

<p>Immobilized embryos were dorsally oriented and the number of mature melanocytes was counted in the defined region of the head. Reduction in melanocyte number was observed in both exon 6- and exon 8-morphants (B and E). Co-injection of wildtype <i>hABCB6</i> mRNA transcript significantly rescued the loss of melanocyte phenotype by increasing the number of mature melanocytes (C and F).</p