Anne-Marie Koop

7 265 Figure 3 c i e g l h k 0 600 cell surface area ( μ m 2 ) sham PAB 200 400 Hand2 f/f MCM-Hand2 f/f * sham PAB fibrosis (%) 0 2 1 3 Hand2 f/f MCM-Hand2 f/f sham PAB 0 relative Col1A expression 4 2 6 8 * Hand2 f/f MCM-Hand2 f/f 10 * d sham PAB 0 relative nppa expression 15 5 25 10 20 * * Hand2 f/f MCM-Hand2 f/f 0 relative nppb expression 1 2 3 4 sham PAB Hand2 f/f MCM-Hand2 f/f sham PAB 0 relative Acta1 expression 0.5 1.0 1.5 * * Hand2 f/f MCM-Hand2 f/f f Hand2 f/f MCM-Hand2 f/f 0 relative Myh7 expression 2 1 3 * sham PAB 0 relative Tgfb expression 2 3 1 Hand2 f/f MCM-Hand2 f/f sham PAB sham PAB 0 relative Eng expression 1 2 Hand2 f/f MCM-Hand2 f/f * sham PAB 0 relative Rcan 1.4 expression 1.5 2.5 0.5 1.0 2.0 ** Hand2 f/f MCM-Hand2 f/f a b 0 3.0 EI ED sham PAB Hand2 f/f MCM-Hand2 f/f 1.5 0.5 2.0 1.0 2.5 * * 0 EI ES sham PAB Hand2 f/f MCM-Hand2 f/f 3.0 1.5 0.5 2.0 1.0 2.5 * * j n m sham PAB 0 relative Vegfr2 expression 1 2 Hand2 f/f MCM-Hand2 f/f 0 2 1 Hand2 f/f MCM-Hand2 f/f sham PAB * * * n o capillaries / cardiomyocyte Figure 3 . Assessment of left ventricular remodelling induced by PAB upon cardiac depletion of Hand2. a) Assessment of left ventricular eccentricity index at end-diastole and b) end-systole; c) Quantification of cardiomyocyte CSA from images of histological sections stained for wheat germ agglutinin (WGA), n=30 microscopic field/heart, 3 hearts. Quantitative real-time PCR analysiswas performed to assess the expression levels of several genes known to be related to cardiac hypertrophy in RV tissue from Hand2F/F and MCM- Hand2F/F animals either after sham or PAB surgery: d) natriuretic peptides atrial natriuretic factor (Nppa); e) brain natriuretic peptide (Nppb); f) β -myosin heavy chain (Myh7); g) α -skeletal actin (Acta1); h) regulator of Calcineurin 1 Isoform 4 (Rcan1-4); i) Quantification of collagen deposition from images of histological sections stained for Sirius Red, n=30microscopic field/heart, 3 hearts. Quantitative real-time PCR analysis of j) collagen type I alpha 1 chain (Col1A1); k) transforming growth factor beta (TGF- β ) and l) endoglin; m) Capillaries in RV sections of Hand2F/F and MCM-Hand2F/F animals subjected to either sham or PAB surgery were identified by isolectin B4 immunohistochemistry combined with WGA and, from the images obtained, we determined the ratio of capillaries per cardiomyocyte ratios; n) Quantitative real-time PCR analysis of vascular endothelial growth factor receptor 2 (Vegfr2) expression levels in the RV of Hand2F/F and MCM-Hand2F/F animals, subjected to either sham or PAB surgery. Data are from 5-10 animals per group; ∗ p < 0.05 between indicated groups (error bars are s.e.m.). Mechanisms underlying pressure overload-induced RV pathological remodelling During RV failure, pathological remodelling is activated, as observed by an increase in cardiomyocyte hypertrophy and fibrosis, followed by a decline in cardiac function. To obtain more insight into the molecular and cellular mechanisms driving RV remodelling induced by pressure overload, we performed total RNA sequencing in RV tissue derived from control (Hand2 F/F ) animals subjected to either sham or PAB surgery. Transcriptomic analysis revealed 1399 genes to be differentially expressed,