AGEs/RAGE Promote Osteogenic Differentiation in Rat Bone Marrow-Derived Endothelial Progenitor Cells via MAPK Signaling

Figure 5

siRAGE transduction reduces the AGE-induced increase in RAGE expression in EPCs and reduces the expression levels of calcification-related proteins in EPCs. (a) Representative western blotting images and densitometric analysis of RAGE protein expression in EPCs stimulated with the indicated concentration of AGE for 7 days. (c) RAGE protein expression levels were detected and semiquantified using western blotting in the EPCs induced by 100 μg/ml AGE for 7 days, after transfection with siRAGE, EV, or treatment with PBS as the control. (e) EPCs were transfected with siRAGE, empty vector (EV-AGEs), or control (EGM-2MV) then induced with 100 μg/ml AGE for 7 days. Cells not treated with AGEs were used as the controls. Expression levels of cellular calcification-related proteins OPG and RUNX2 were detected by western blotting. GAPDH was used as the loading control. Each data point represents the results of three independent assays. Comparison with control, ^▲ vs.; AGEs 100 mg/ml compared with other groups; ^◆ vs.; comparison with AGEs alone, vs.; comparison with EV-AGEs, ^# vs. AGE: advanced glycation end product; EPC: endothelial progenitor cell; EV: empty-vector–transfected control group; OPG: osteoprotegerin; RUNX2: runt-related transcription factor 2; siRAGE: small interfering RNA against receptor for AGE.

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