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

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

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

Characteristics of the study population.

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