Supplementary Material
Supplementary Figures
Supplementary Figure 1. LV function remains similar between baseline, cRVH and dRVH animals.
(A) LV+Septum weight remains similar between baseline, cRVH and dRVH animals (n=6 animals for baseline, 10 for cRVH and 9 for dRVH).
(B) LV function assessed by fractional shortening (FS), ejection fraction (EF) and mass weight remains similar between baseline and dRVH animals. Representative parasternal short axis image of the LV in m-mode is shown. Septal wall is shown at the top and LV free wall is shown below, all measurements were made through the papillary muscle to keep consistency of measurements between animals.
Supplementary Figure 2. Ponseau-S confirms loading control of actin for all performed immunoblots.
Ponseau-S has a very similar trend to the observed actin shown in Figure 2C, suggesting that actin is an appropriate loading control for all performed immunoblots.
Supplementary Figure 3. HIF1mRNA levels are decreased in dRVH compared to cRVH.
(A) mRNA levels of HIF1, measured by qRT-PCR, in the3 groups of rats are shown (n=5 animals per group, *p<0.05 vs. baseline, #p<0.05 vs. cRVH).
(B) Nuclear levels of HIF1 (HIF1: red, nuclear stain DAPI: blue) and cytoplasmic levels of Glut 1 (green) are similar between the RVs from control and dRVH animals as indicated by confocal microscopy and immunofluorescence. Note the absence of nuclear HIF1 and cytoplasmic Glut1 levels in these two groups of RVs compared to the cRVH group shown in Figure 3B.
Supplementary Figure 4. Lectin quantification for assessment of capillaries.
Representative image (left) and high magnification (right circle) showing the quantification of lectin (green) using a methodology described in [13]. A grid consisting of 2070 evenly distributed points (in red) was overlaid on each lectin image (representing stacked images of a standardized volume of RV tissue as described in the methods). Then the total number of points that are associated with capillaries (marked by lectin) are divided by the total number of points associated with the RV sample in each image; the volume of RV samples studied was kept identical among all the RVs (n=5-10 animals/group, as also shown in Figure 6B; *p<0.05 vs. baseline, #p<0.05 vs. cRVH).
Supplementary Figure 5. Apoptosis in cRVH and dRVH.
dRVH and cRVH myocardiumhasdetectableapoptosis measured by TUNEL staining (TUNEL: green, nuclear stain DAPI: blue) measured by confocal microscopy and immunofluorescence. Representative confocal images magnification (left) and quantified mean data (right) are shown (n=5 images per animal and 5 animals/group).
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