Figure 1

BMSCs infection and verification of secreted sFlt-1. After the BMSCs were confirmed by flow cytometric analysis, BMSCs were grown to 90% confluence and infected with adenoviruses at a multiple of infection of 3000 for 2 hours. 24 hours later, BMSCs were harvested and ready for use. Fluorescence microscope showed up to 100% GFP-positive cells. (Figure 1-A). Supernatants deposits from sFlt-1 bearing BMSCs could be recognized by antibodies reactive to NH₂ terminus of mouse Flt-1, but negative staining in supernatants deposits from control BMSCs in Western blot analysis. (Figure 1-B). Conditioned media from Adv-sFlt-1 infected BMSCs was shown to inhibit the VEGF-driven mouse endothelial cells proliferation by about 50% compared with controls (Figure 1-C, P < 0.05).