Structure of <i>NLRP6/AVR</i> and <i>ADM</i> genes and location of the SNPs analyzed.

<p>Exons (shown as boxes) 1–8 for <i>NLRP6/AVR</i> and exons 1–4 for <i>ADM</i> are shown. Gene untranslated (5′-untranslated and 3′-untranslated) regions are unfilled. Corresponding nucleotide positions for the <i>NLRP6/AVR</i> and <i>ADM</i> loci on chromosome 11 are indicated in bp. Locations of the SNPs genotyped are shown by vertical lines. </p

Identification of the 12T-ins/del polymorphism in the <i>ATP1A1</i> promoter region and transcriptional activity of <i>ATP1A1</i> promoter variants.

<p>(A) Nucleotide sequence spanning the poly-T sequence involved in the 12T-ins/del polymorphism from the two <i>ATP1A1</i> (p12T ins, p12T del) reporter gene constructs utilized in the transcriptional assays. On right detection of 12T-insertion and 12T-deletion alleles by PCR-amplification followed by denaturing polyacrylamide gel (6%) electrophoresis used for genotyping of the Sardinian cohort. (B) Illustration of the <i>ATP1A1</i> promoter region. Non-coding exon is presented as open box and exon encoding the NH₂-terminal region is presented as black box. Sequence and location of the <i>ATP1A1</i> 12T-ins/del polymorphism is shown. The positions of TATAAA box, INR (initiator) and DPE (downstream promoter element) core promoter elements within <i>ATP1A1</i> promoter are shown. Core promoter elements were identified based on 100% homology with corresponding consensus sequences [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0116724#pone.0116724.ref039" target="_blank">39</a>,<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0116724#pone.0116724.ref040" target="_blank">40</a>]. (C) Schematic of two <i>ATP1A1</i> (p12T ins, p12T del) reporter gene constructs. (D) Relative transcriptional activity of 12T-insertion (p12T ins) and 12T-deletion (p12T del) gene constructs in Cos1, HEK293 and MDA-MB-468 cells. *, <i>P</i> < 0.05; **, <i>P</i> < 0.01 (two-tailed student <i>t</i>-test).</p

Analysis of ATP1A1 protein expression, blood pressure, heart rate and activity in heterozygous <i>ATP1A1^+/−</i> and wild-type male mice.

<p>(A) Western blot analysis of <i>ATP1A1^+/−</i> and wild type mouse whole kidney extracts (30 μg) reacted with anti-mouse ATP1A1 and anti-mouse βActin polypeptides. (B) Densitometry analysis of samples shown in (A) detecting 58% decrease ATP1A1 levels in <i>ATP1A1^+/−</i> kidneys. (C) Mean systolic blood pressure ± sem (SBP; mmHg). (D) Mean heart rate ± sem (beats/min; BPM). (E) Mean activity ± sem (Counts/min) in <i>ATP1A1^+/−</i> (solid bars, n = 4) and wild-type (open bars, n = 5) male mice. * <i>P</i> < 0.03, ** <i>P</i> = 0.015 (two-tailed student <i>t</i>-test).</p

Characteristics of the study population.

<p>Characteristics of the study population.</p

Analysis of <i>ATP1A1</i> (12T-ins/del) variants based on blood pressure as a quantitative trait.

<p>Blood pressures were adjusted for age, body mass index and case/control status.</p><p>^a Number of individuals</p><p>^b Systolic blood pressure in mmHg</p><p>^c Diastolic blood pressure in mmHg</p><p>s.e.m., standard error of the mean</p><p>Δ SBP, difference in systolic blood pressure</p><p>Δ DBP, difference in diastolic blood pressure</p><p><i>P</i>, Mann-Whitney Rank Sum Test <i>P</i> values.</p><p>Analysis of <i>ATP1A1</i> (12T-ins/del) variants based on blood pressure as a quantitative trait.</p

Mean inbreeding coefficients.

<p>Mean inbreeding coefficients, standard errors (SE) and T test <i>p-Values</i> of Sardinia macro-areas and peninsular Italy.</p

Nine genomic regions showing signals of positive selection in the Sardinian's genome ordered by |iHS|.

<p>Column headers: Position on NCBI36/hg18 of region showing evidence for selection; Size in Kb of the genomic region; nSNP |iHS|>4 indicates the number of SNPs with an absolute |iHS| higher than 4 in each region; nSNP is the number of SNPs in each region; Max iHS is the highest value of each region; Max iHS SNP is the polymorphism with the highest value for each region; P-values: nominal p-values; Empirical p-values: after permutation-based multiple testing corrections; Genes: the genes within the region. When, in the genomic region, there are more than 4 genes, only the first 4 are indicated.</p

ADMIXTURE software results for K = 2–4.

<p>Ancestry for each individual inferred using ADMIXTURE software.</p

Map of Mediterranean basin showing the localization of Sardinia and Sardinian linguistic domains.

<p>A) Map of the Mediterranean basin showing the geographic position of Sardinia. B) The Sardinian linguistic domains: 1 = <i>Gallurese</i> (77 individuals); 2 = <i>Nuorese</i> (88); 3 = <i>Logudorese</i> (385); 4 = <i>Sassarese</i> (342); 5 = <i>Alghero</i> (87); 6 = <i>Campidanese</i> (98).</p

Mean genomic inbreeding coefficients (F_RoH %) using 0.5 and 5 Mb minimum RoH thresholds and mean sum of RoH.

<p>* <i>p-value</i> smaller than 0.05 when comparing each linguistic macro-area to peninsular Italy.</p