Effects of harmine on hepatic SAHH and NQO1 activities, and on plasma Hcy levels in Tg 189N3 mice.

<p>Hepatic SAHH (A) and NQO1 (B) activities are presented as percent of untreated (Vehicle) non-transgenic (Tg –) mice activities. (C) Plasma Hcy level. Data correspond to means ± SEM and the statistical analysis was done with one-way ANOVA followed by Student's unpaired <i>t</i>-test. n = number of mice.</p

DYRK1A protein expression in liver of CBS-deficient mice crossbred with 152F7 transgenic mice.

<p>(A) Western immunoblots showing DYRK1A expression in liver of wild type mice (<i>Cbs</i>^+/+ Tg -), heterozygous mice (<i>Cbs</i>^+/− Tg -), 152F7 transgenic mice (<i>Cbs</i>^+/+ Tg 152F7), and heterozygous mice crossbred with 152F7 transgenic mice (<i>Cbs</i>^+/− Tg 152F7). Proteins were subjected to immunoblot analysis using antibodies specific to DYRK1A (85.5 kDa). After stripping, the membranes were reprobed with anti-β-actin antibody (41.7 kDa) for the control. (B) Relative protein expression was determined by normalization of the density of images from DYRK1A with that of β-actin of the same blot. The values of <i>Cbs</i>^+/− Tg -, <i>Cbs</i>^+/+ Tg 152F7, or <i>Cbs</i>^+/− Tg 152F7 were normalized to the mean of <i>Cbs</i>^+/+ Tg - mice. The blots are representative of three independent experiments. Data correspond to means ± SEM and the statistical analysis was done with one-way ANOVA followed by Student's unpaired <i>t</i>-tests. n = number of mice.</p

Relative expression of DYRK1A, SAHH and NQO1 and SAHH and NQO1 activities obtained from lymphoblastoid cell lines.

<p>The values of lymphoblastoid cell lines (LCLs) from patients with DS (T21) were normalized to the mean lymphoblastoid cell lines from control individuals (control). Data correspond to means ± SEM and the statistical analysis was done by Student's unpaired <i>t</i>-tests. n = number of LCLs. * <i>p</i><0.05; <b>^†</b><i>p</i><0.006.</p

Relative expression of SAHH, BHMT, MS and MTHFR gene based upon Q-PCR data obtained from non-transgenic and Tg189N3 mice.

<p>The values of Tg189N3 were normalized to the mean Tg – mice. Data correspond to means ± SEM and the statistical analysis was done by Student's unpaired <i>t</i>-tests. n = number of mice. *<i>p</i> = 0.068; <b>^†</b><i>p</i>< 0.03.</p

Hepatic DYRK1A mRNA and protein expression in liver of transgenic mice.

<p>Relative expression of DYRK1A gene was based on Q-PCR data and protein expression was determined by normalization of the density of images from DYRK1A with that of β-actin of the same blot. The values of Tg 152F7, Tg189N3 and Ts65Dn were normalized to the mean Tg – mice from each lines. The blots are representative of three independent experiments. Data correspond to means ± SEM and the statistical analysis was done by Student's unpaired <i>t</i>-tests. n = number of mice.</p

Hepatic SAHH activity is increased in Tg 189N3 and Ts65Dn transgenic mice.

<p>SAHH activity in (A) Tg 189N3 and (B) Ts65Dn transgenic mice. The values were normalized to the mean of Tg – mice from each line. Data correspond to means ± SEM and the statistical analysis was done by Student's unpaired <i>t</i>-test. n = number of mice.</p

Primer sequences for real-time quantitative reverse transcription-polymerase chain reaction (Q-PCR) (all primers are listed 5′ to 3′).

<p>Primer sequences for real-time quantitative reverse transcription-polymerase chain reaction (Q-PCR) (all primers are listed 5′ to 3′).</p

Plasma Hcy level is decreased in 152F7 transgenic mice and in CBS-deficient mice crossbred with 152F7 transgenic mice.

<p>(A) Plasma Hcy level, (B) hepatic CBS, (C) SAHH and (D) NQO1 activity in wild type mice (<i>Cbs</i>^+/+ Tg -), heterozygous mice (<i>Cbs</i>^+/− Tg -), 152F7 transgenic mice (<i>Cbs</i>^+/+ Tg 152F7), and heterozygous mice crossbred with 152F7 transgenic mice (<i>Cbs</i>^+/− Tg 152F7). The values were normalized to the mean of <i>Cbs</i>^+/+ Tg - mice. Data correspond to means ± SEM and the statistical analysis was done with one-way ANOVA followed by Student's unpaired <i>t</i>-tests. n = number of mice.</p

Classification of heart defects (CHD+) according to the distribution frequency of their p-values.

<p>Empirical cumulative distributions of p-values for the 2-by-2 Student tests A: AVSD/ASD; B: AVSD/VSD; C: ASD/VSD. Comparisons were obtained for 11 224 tests. The comparison to the uniform distribution shows that there is no overall differential expression between ASD and VSD (less small p-values than for an uniform distribution). Cdf: cumulative distribution function.</p

Transcriptome signature of heart defects.

<p>Comparison of ratios for DE genes up- or down-regulated in the AVSD or ASD+VSD group between (1) array ratio in the Ts21 CHD⁻ vs 2N, (2) array ratio in CHD⁺ vs CHD⁻, (3) q-PCR ratio from different experiments in CHD⁺ vs CHD⁻ using additional samples (total number in parentheses). Gene nomenclature, <i>AUTS2</i>: autism susceptibility candidate 2; <i>CNN2</i>: calponin 2; <i>DSCR3</i>: Down syndrome critical region gene 3; <i>DYNLT3</i>: dynein, light chain, Tctex-type 3; <i>NQO1</i>: NAD(P)H dehydrogenase, quinone 1; <i>OFD1</i>: oral-facial-digital syndrome 1; <i>PDIA4</i>: protein disulfide isomerase family A, member 4; <i>PIGP</i>: phosphatidylinositol glycan anchor biosynthesis, class P; <i>TTC3</i>: tetratricopeptide repeat domain 3; <i>TUBB2B</i>: tubulin, beta 2B; <i>ACTG1</i>: actin, gamma 1; <i>ALDOC</i>: aldolase C, fructose-bisphosphate; <i>CACYBP</i>: calcyclin binding protein; <i>DTYMK</i>: deoxythymidylate kinase (thymidylate kinase); <i>ENO2</i>: enolase 2 (gamma, neuronal); <i>GART</i>: phosphoribosylglycinamide formyltransferase, phosphoribosylglycinamide synthetase, phosphoribosylaminoimidazole synthetase; <i>IFNAR1</i>: interferon (alpha, beta, and omega) receptor 1; <i>PLTP</i>: phospholipid transfer protein; <i>RPL10A</i>: ribosomal protein L10a; <i>TNFAIP2</i>: tumor necrosis factor, alpha-induced protein 2.</p