Figure 3

Effects of AGEs on late EPC migration, adhesion and SDF-1/CXCR4 system. Late EPCs were treated with different concentrations of AGEs (0, 50, 100, 200, and 500 μg/ml) for 24 h. A: The modified Boyden chamber assay was used with VEGF as a chemoattractive factor for late EPC migratory function. The migrated cells were stained with DAPI and counted under microscope. B: EPCs with equal cell numbers were seeded on fibronectin coated culture dishes and incubated for 30 min at 37°C. Adherent cells were counted for late EPC adhesion function. C: The supernatants were collected, and the content of SDF-1 was analyzed by ELISA. D: The CXCR4 mRNA expressions were determined using real time quantitative RT-PCR. E: Cell lysates were resolved on 12% SDS-PAGE, followed by transfer to PVDF membrane. Western blot was carried out with specific antibody for CXCR4, and each band was detected by the ECL reagent. In addition, the β-actin was analyzed as loading control and protein. Data represent the mean ± SE of four different experiments. *p < 0.05, **p < 0.01 vs. control.