Figure 5
Forced NICD expression induces multilineage differentiation of total and c-KitPOS/NKX2.5POS bone marrow mesenchymal stem cells. Total bone marrow mesenchymal stem cells (BMSCs) and c-KitPOS/NKX2.5POS BMSCs were infected with NICD-Ad or NC-Ad, with uninfected cells serving as an additional control (MOCK). After 8 days, the cells were harvested for (A) real-time quantitative RT-PCR analysis and (B) immunofluorescence staining for markers for cardiomyocytes (NKX2.5 and cardiac troponin T (cTnT)), smooth muscle cells (SM22α), and endothelial cells (von Willebrand factor (vWF)). Triplicate experiments were performed for real-time quantitative RT-PCR analysis (n = 3). *P <0.05, ***P <0.001, # P <0.05, ## P <0.01, ### P <0.001 versus other associated groups. For immunofluorescence staining, images were captured by confocal microscopy. Target proteins were detected with phycoerythrin-conjugated or fluorescein isothiocyanate-conjugated IgG. Nuclei were counterstained with 4′,6-diamidino-2-phenylindole (DAPI). Scale bar: 50 μm. EC, endothelial cell; NC-Ad, negative control-expressing adenovirus; NICD, notch receptor intracellular domain; NICD-Ad, NICD-expressing adenovirus; SMC, smooth muscle cell.