Fig. 3.

V0 and V1 are the main isoforms expressed during fibrosis development during pressure overload. (A-D) mRNA expression of versican isoforms V0, V1, V2 and V3 from the left ventricle of mouse hearts at day 1, 3, 7, 14, 28 and 56 after aortic banding (AB, n = 8–11) or sham operations (Control, n = 9–10). (E-G) Representative western blot image and quantification (normalized to total protein stain, relative to day 1 post AB) of the V0 and V1 isoform from the left ventricles of mice at day 1, 3, 7, 14, 28 and 56 after aortic banding (AB, n = 8) or sham operations (Control, n = 5–8). All samples were treated with chondroitinase ABC enzyme to remove glycosaminoglycan chains, except for the sample on the right side of the image. Pooled extracellular matrix fractions from left ventricular tissue of aortic banded mice obtained after 8 to 10 weeks were used as positive controls. Control samples (sham operation) displayed very low or no amount of versican. The absolute quantity of mRNA expression (copies/μl measured by ddPCR) relative to mRNA expression measured by primer pairs targeting all isoforms of Vcan is presented. Data represent mean ± SD. Repeated measures two-way ANOVA (A-D) or one-way ANOVA (F) with Bonferroni’s multiple comparisons test, and Kruskal-Wallis (G) with Dunn’s post-hoc test were used for statistical analysis. P values < 0.05 were considered statistically significant. *P < 0.05 Control vs. AB, ^†P < 0.05 AB vs. day 1 post AB, ^‡P < 0.05 AB vs. day 3 post AB. Note the differences in the scaling of y-axis of the line graphs (A-D and F-G).