Figure 5.

Co-culture of cardiac fibroblasts with mesenchymal stem cells and the direct effects of pigment epithelium-derived factor on cardiac fibroblasts. (A) Bar graph illustrates proliferation assays of cardiac fibroblasts co-cultured with mesenchymal stem cells. Cardiac fibroblasts co-cultured with cardiac fibroblasts served as control. (B) Bar graph illustrates migration assays of cardiac fibroblasts co-cultured with mesenchymal stem cells. Cardiac fibroblasts co-cultured with cardiac fibroblasts served as control. (C) Bar graph shows the direct effect of pigment epithelium-derived factor on cardiac fibroblasts proliferation. (D) Representative flow cytometry analyses for cell cycle and the bar graph shows the effect of pigment epithelium-derived factor on cardiac fibroblasts' cell cycle. (E) Representative western blotting analyses and bar graphs demonstrate that pigment epithelium-derived factor regulated cardiac fibroblasts expression of cyclin D1 and P27. (F) The effect of pigment epithelium-derived factor on cardiac fibroblasts apoptosis. Representative Hoechest stained images and bar graphs illustrating degree of apoptosis for cardiac fibroblasts analysed by Hoechest staining or flow cytometry. White arrows show representative apoptotic cells with nuclear fragmentation. (G) The effect of pigment epithelium-derived factor on cardiac fibroblast migration. Representative crystal violet staining of migrated cardiac fibroblasts in the lower compartments of the transwell system and bar graph illustrating the effect of pigment epithelium-derived factor on cardiac fibroblasts migration in transwell system. All data expressed as means ± SEM. n = 6 per group. *P< 0.05, **P< 0.01, ***P≤ 0.001.