Position and stem penalties for pathogenicity prediction.

<p>(A) The variant history and conservation scores at the analogous positions of every mitochondrial tRNA were averaged and used to score a generic tRNA structure. This identifies the regions of the tRNA that are most vulnerable to pathogenic variants. (B) Variants at base pairing regions are assessed based on the steric hindrance they induce, with the highest scores assigned at the ends of the stems region as shown in the scoring heat map for the phenylalanine tRNA.</p

Separation of benign and pathogenic variants by MitoTIP.

<p>(A) Pathogenicity scores from naïve evaluations of known pathogenic (n = 38) and described benign (n = 651) variants plotted using box and whiskers at 5–95% (<i>p</i><0.0001 by Mann Whitney test). Negative scoring is possible when polymorphisms improve Watson-Crick pairing in stems. (B) Sensitivity and specificity plot of these data at a range of pathogenicity scores. The crossover pathogenicity score was 12.8.</p

Comparison between predictive systems.

<p>Comparison between predictive systems.</p

Phenylephrine-mediated contraction in mesenteric arteries submitted to high blood flow.

<p>Phenylephrine-mediated contraction was measured in mesenteric arteries submitted to a chronic increase in blood flow (high flow: HF, right panel) and in control arteries submitted to normal flow (NF, left panel). Arteries were isolated from ovariectomized rats treated with resveratrol 5.0 (R5.0 or Resv. 5.0, n = 9 rats) or 37.5mg/kg (R37.5 or Resv. 37.5, n = 9 rats) or with the vehicle (n = 10 rats). Mean ± sem is represented. *P<0.05, HF versus NF arteries. ^#P<0.05, R37.5 versus vehicle.</p

Arterial diameter in mesenteric arteries submitted to high blood flow.

<p>Luminal diameter was measured in mesenteric arteries submitted to a chronic increase in blood flow (high flow: HF) and in control arteries submitted to normal flow (NF). Arteries were isolated from ovariectomized rats treated with resveratrol 5.0 (B, n = 9 rats) or 37.5mg/kg (C, n = 9 rats) or with the vehicle (A, n = 10 rats). In a separate series of experiments, ERalpha-/- mice were ovariectomized and treated with the solvent or with resveratrol 5mg/kg (D, n = 4 mice per group). Mean ± sem is represented. *P<0.05, HF versus NF arteries.</p

RSV increases the expression of anti-oxidant enzymes in brain mitochondria.

<p>Expression of MnSOD was analyzed by Western blotting (normalised to the quantity of VDAC protein) on mitochondrial fractions from young (Y) and old (O) control (Ctl) and RSV-treated mice. (A) Representative blot of two duplicate experiments on n = 4 animals. (B) Quantification of MnSOD expression. Data are represented as means ± <i>sem</i> of four animals in duplicate. The (+) indicates significant differences (p<0.05) between young and old animals and the (*) shows a significant effect of the RSV-diet (p<0.05).</p

Mitochondrial proteins expression level.

<p>The expression level of of Cytochrome-C Oxidase IV (COX IV, A), sirtuin-1 (B), pGC1alpha (C), and Cytochrome C (D) was determined using Western-blot analysis in mesenteric arteries (left panel). The ratio of the expression level in HF to the expression level in NF arteries is shown in the right panel. Arteries were isolated from ovariectomized rats treated with resveratrol 5.0 (Resv. 5.0, n = 9 rats) or 37.5mg/kg (Resv. 37.5, n = 9 rats) or with the vehicle (n = 10 rats). Mean ± sem is represented (n = 12 rats per group). Blots are shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0146148#pone.0146148.s003" target="_blank">S3A–S3D Fig</a>. *P<0.05, effect of the treatment: Resv. 5.0 or Resv. 37.5 versus vehicle.</p

Expression level of eNOS (A), p67 phox (B), Gp91 phox (C) and ERK1/2 activation (D, ratio of phospho-ERK1/2 to ERK1/2) expression level.

<p>The ratio of protein expression in HF to the expression level in NF arteries is shown in the right panel. Arteries were isolated from ovariectomized rats treated with resveratrol 5.0 (Resv. 5.0, n = 9 rats) or 37.5mg/kg (Resv. 37.5, n = 9 rats) or with the vehicle (n = 10 rats). Mean ± sem is represented. Blots are shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0146148#pone.0146148.s003" target="_blank">S3A–S3D Fig</a>. *P<0.05, effect of the treatment: Resv. 5.0 or Resv. 37.5 versus vehicle.</p

RSV specifically targets Complex I activity in brain mitochondria.

<p><b>(A) Maximal activity of complexes in mitochondria incubated with 5</b> μ<b>M RSV. (B) Maximal activity of complexes in mitochondria incubated with 50</b> μ<b>M RSV.</b> Mouse brain mitochondria disrupted by frozen/thawed cycles were incubated with either vehicle (DMSO 1/2000) or RSV (50 μM) just prior analyzing complex I to IV activities. Data are expressed as the percentage of complex activities in vehicle treated mitochondria and are represented as mean <i>± sem</i> of n = 5 animals in duplicate. The (*) indicates significant differences (p<0.05) between vehicle and RSV treated mitochondria.</p

RSV increases the NFR and NUR activities but decreases the NCCR activity.

<p>Control young mice brain mitochondria were incubated with either DMSO (1/2000, vehicle) or 5 μM RSV (30 minutes) and NADH Ubiquinone Reductase (NUR), NADH:FeCN reductase (NFR), and NADH cytochrome C reductase (NCCR) activities were measured on incubated mitochondria. Data are represented as mean <i>± sem</i> of n = 3 animals in duplicate. The (*) indicates significant differences (p<0.05) between vehicle and RSV treated mitochondria.</p