Sanger sequencing confirmation of the disease segregating variant identified in the linked region.

<p>Candidate region identified using linkage analysis in PACG pedigree animals (left). Sequence chromatograms from a carrier (C), confirms heterozygous genotype whereas affected animals (A) display a homozygous state of variant identified in <i>NEB</i> (g.5588214 A->G) (Right).</p

Whole genome homozygosity mapping results.

<p>A red peak reaching a 1.0 max score of statistical significance depicts a homozygous region on chromosome 19. The region coincides with the 0.49 Mbp locus identified using two-point linkage analysis and is bracketed by recombination spots (indicated in red). The identified haploblock fulfills the zygosity criterion by displaying homozygosity in affected animals (indicated in red) and heterozygosity in unaffected carriers (indicated in green).</p

Basset Hound Pedigree used in this study.

<p>The affected Basset 5a in the second generation was duplicated twice (5b and 5c) in order to break two otherwise computationally confounding breeding loops. Genotypes of typed markers within region uncovered following two-point linkage analysis are shown. Shading indicates the transmission pattern of heterozygous parental haplotypes to affected, homozygous offspring. Additional patterns of shading including diagonal lines indicate variation to the same haplotype identified in the other pedigree members. Complete concordance of homozygous haplotype inheritance with the disease phenotype is observed in all affected animals.</p

Genome-wide linkage analysis of SNP genotype data.

<p>Using two-point linkage analysis, a maximum LOD_two-point score of 3.07 was achieved for a 0.49 Mb locus on chromosome 19. The red line of statistical significance indicates LOD scores values > 3 (A). Using multipoint linkage analysis, an increased maximum LOD_multipoint score of 3.24 was achieved for a locus mapped to the same location (B). A schematic view of the maximum LOD scores achieved across chromosome 19. The statistically significant locus is located at the distal end of chromosome 19 (Chr19: 54,949,124–56,765,346) and spans 1.82 Mbp (C).</p

A Fisher exact contingency table of genotypes observed in a confirmatory animal cohort for the <i>RIF1</i> variant (g.55723957 C->T).

<p>Forty-four additional unaffected and affected Basset Hounds were selected for confirmatory sequencing of the <i>RIF1</i> variant (g.55723957 C->T). The observed total of individuals and percentage of individuals displaying a specific genotype is shown for each cell respectively. The two-tailed P value is 0.57 (Fisher Exact Probability Test)</p><p>A Fisher exact contingency table of genotypes observed in a confirmatory animal cohort for the <i>RIF1</i> variant (g.55723957 C->T).</p

A Fisher exact contingency table of genotypes observed in a confirmatory animal cohort for the <i>NEB</i> variant (g.55885214 A->G).

<p>Forty-four additional unaffected and affected Basset Hounds were selected for confirmatory sequencing of the <i>NEB</i> variant (g.55885214 A->G). The observed total of individuals and percentage of individuals displaying a specific genotype is shown for each cell respectively.</p><p>The two-tailed P value is 0.00034 (Fisher Exact Probability Test)</p><p>A Fisher exact contingency table of genotypes observed in a confirmatory animal cohort for the <i>NEB</i> variant (g.55885214 A->G).</p

Alignment of the amino acid sequence of exon 48 in Nebulin in several vertebrate species.

<p>The Lysine (K) residue (highlighted in blue) at position p.2051 is conserved among 23 vertebrate species.</p

Recurrent missense variants identified in two of 289 candidate genes in 12 sporadic CD subtype of AMD cases by WES.

<p>Recurrent missense variants identified in two of 289 candidate genes in 12 sporadic CD subtype of AMD cases by WES.</p

Segregation analysis of rare sequence variants identified in candidate genes in cuticular drusen (CD) families by whole exome sequencing (WES).

<p>Circles, females; squares, males; empty symbols, unaffected; black symbols, affected; asterisks, exome sequenced individuals; ‘+’ symbol, wild type allele; ‘m’ symbol, mutant type allele. The age at participation is specified below the symbols.</p

Pedigrees of six cuticular drusen (CD) families in which whole exome sequencing (WES) was performed.

<p>Circle and square symbols indicate female and male individuals, respectively. Symbols with slashes indicate deceased individuals. Black and empty symbols indicate affected and unaffected individuals, respectively. Asterisks indicate the family members for whom exome sequencing was performed. The numbers below the symbols indicate the age at participation of family members.</p