Representative Sirius red stained cross-sections of sham C57BL/6 <i>Ldlr</i>^−/−<i>Cbs</i>^+/− mice, control MI mice, and AdCBS MI mice at day 28 after ligation of the LAD.

<p>Morphometric analysis was performed on tissue sections of 4 separate regions using Axiovision 4.6 software (Zeiss). Scale bar represents 1 mm.</p

(A) Relative protein expression levels of CBS in the liver of C57BL/6 <i>Ldlr^−/− Cbs^+/+</i> mice (grey bar) and of C57BL/6 <i>Ldlr^−/− Cbs^+/−</i> mice 2 weeks after gene transfer with 10¹¹ particles of Adnull (black bar) or 10¹¹ particles of AdCBS (black crosshatched bar).

<p>Data in Adnull- and AdCBS-treated mice were calculated using values in C57BL/6 <i>Ldlr^−/− Cbs^+/+</i> mice as the denominator. All data are shown as means ± SEM (n = 7 to 9 for each condition). (B) Representative Western blot of CBS expression in the liver of C57BL/6 <i>Ldlr^−/− Cbs^+/+</i> mice (lanes 1 and 2) and of C57BL/6 <i>Ldlr^−/− Cbs^+/−</i> mice injected with 10¹¹ particles of Adnull (lanes 3–5) or the same dose of AdCBS (lanes 6–8).</p

Hemodynamic parameters in the left ventricle and in the aorta of sham C57BL/6 mice and of sham C57BL/6 <i>Ldlr</i>^−/−<i>Cbs</i>^+/− and sham AdCBS treated C57BL/6 <i>Ldlr</i>^−/−<i>Cbs</i>^+/− mice.

<p>Sham C57BL/6 mice (n = 10) were fed normal chow. Sham C57BL/6 <i>Ldlr</i>^−/−<i>Cbs</i>^+/− (n = 11) and AdCBS treated (n = 11) C57BL/6 <i>Ldlr</i>^−/−<i>Cbs</i>^+/− mice were fed with folate-depleted, methionine-enriched diet supplemented with 0.2% cholesterol and 10% coconut oil. Data are expressed as means ± S.E.M.</p

Morphometric analysis of the left ventricle of sham C57BL/6 mice and of sham C57BL/6 <i>Ldlr</i>^−/−<i>Cbs</i>^+/− and sham AdCBS treated C57BL/6 <i>Ldlr</i>^−/−<i>Cbs</i>^+/− mice.

<p>Sham C57BL/6 mice (n = 10) were fed normal chow. Sham C57BL/6 <i>Ldlr</i>^−/−<i>Cbs</i>^+/− (n = 11) and sham AdCBS treated (n = 11) C57BL/6 <i>Ldlr</i>^−/−<i>Cbs</i>^+/− mice were fed with folate-depleted, methionine-enriched diet supplemented with 0.2% cholesterol and 10% coconut oil. Data are expressed as means ± S.E.M.</p><p>LV: left ventricular.</p

Histological parameters of the left ventricular myocardium of Sham C57BL/6 <i>Ldlr</i>^−/−<i>Cbs</i>^+/− mice and of the infarct area and the remote myocardium at day 28 after ligation of the LAD in control and AdCBS treated C57BL/6 <i>Ldlr</i>^−/−<i>Cbs</i>^+/− mice.

<p>Data are expressed as means ± S.E.M. (n = 11 for Sham, n = 13 for Control MI, n = 25 for AdCBS MI). N.A.: not applicable.</p>§<p>p<0.05;</p>§§<p>p<0.01;</p>§§§<p>p<0.001 versus Sham.</p>*<p>p<0.05 versus Control MI.</p

(Immuno)histochemical analysis of the infarct area in control MI and AdCBS MI mice at day 28 after ligation of the LAD.

<p>Representative photomicrographs show CD31 positive capillaries and Sirius red stained collagen viewed under polarized light. Scale bar represents 50 µm.</p

Heart and lung weights of sham C57BL/6 mice and of sham C57BL/6 <i>Ldlr</i>^−/−<i>Cbs</i>^+/− and sham AdCBS treated C57BL/6 <i>Ldlr</i>^−/−<i>Cbs</i>^+/− mice.

<p>Sham C57BL/6 mice (n = 10) were fed normal chow. Sham C57BL/6 <i>Ldlr</i>^−/−<i>Cbs</i>^+/− (n = 11) and sham AdCBS treated (n = 11) C57BL/6 <i>Ldlr</i>^−/−<i>Cbs</i>^+/− mice were fed with folate-depleted, methionine-enriched diet supplemented with 0.2% cholesterol and 10% coconut oil. Data are expressed as means ± S.E.M.</p

Time course of plasma homocysteine levels (A) and plasma cholesterol (closed symbols) and HDL cholesterol levels (open symbols) (B) in female C57BL/6 <i>Ldlr</i>^−/−<i>Cbs</i>^+/− control mice or in female C57BL/6 <i>Ldlr</i>^−/−<i>Cbs</i>^+/− mice injected with 5×10¹⁰ particles of AdCBS.

<p>A hyperhomocysteinemic and high saturated fat/high cholesterol diet (0.2 mg/kg folic acid, 4.1 g/kg L-methionine, 1.25% cholesterol (w/w), and 10% coconut oil (v/w)) was initiated 3 weeks before adenoviral gene transfer or saline injection and maintained throughout the experiment. The 0 week time point corresponds to the time point of gene transfer in the intervention group. Data are expressed as means ± S.E.M. (n = 8).</p

Hemodynamic parameters in the left ventricle and in the aorta of Sham C57BL/6 <i>Ldlr</i>^−/−<i>Cbs</i>^+/− mice and <i>at</i> day 28 after myocardial infarction in control and AdCBS treated C57BL/6 <i>Ldlr</i>^−/−<i>Cbs</i>^+/− mice.

<p>Data are expressed as means ± S.E.M.</p>§<p>p<0.05;</p>§§§<p>p<0.001 versus Sham.</p>*<p>p<0.05 versus Control MI.</p

Heart and lung weights in sham C57BL/6 <i>Ldlr</i>^−/−<i>Cbs</i>^+/− mice and at 28 days after myocardial infarction in control and AdCBS treated C57BL/6 <i>Ldlr</i>^−/−<i>Cbs</i>^+/− mice.

<p>Data are expressed as means ± S.E.M. (n = 11 for Control reference, n = 13 for Control MI, n = 25 for AdCBS MI).</p>§<p>p<0.05;</p>§§§<p>p<0.001 versus Sham.</p>*<p>p<0.05 versus Control MI.</p