Figure 4

Effects of diabetes on structural remodeling and protein levels in the Wt and Hif1a ^+/- LV. (A) Representative of immunofluorescence confocal and light microscopy images of the hearts stained with anti-connexin 43 (Cx43, red) together with fluorescein-labeled wheat germ agglutinin (WGA, green); with picrosirius red (PSR, polarizing microscopy); and with anti-collagen 1 (Col1, red), autofluorescence (auto) are shown. The nuclei were counterstained with Hoechst 33342 (blue). Confocal images are stacked Z-plane sections from confocal microscopy. Scale bar: Cx43/WGA 25 μm, PSR 50 μm, Col1 25 μm. (B) Representative Western blot analyses of protein lysates from the isolated LVs of Wt and Hif1a ^+/- non-diabetic and diabetic hearts are shown. (C-E) A relative quantification of protein levels of Cx43, phosphoCx43 (pCx43), and Col1 (n = 3 per group) was performed. The bar graphs show the mean values of relative protein levels normalized to the loading control (GAPDH) ± SEM from densitometric analysis. *P < 0.05 (2-way ANOVA with Tukey’s post-test).