Figure 1

Effects of GLAP on ROS generation (A) and RAGE gene expression in HUVECs (B), secondary structure of GLAP-aptamer (C), and binding affinity of GLAP to immobilized vRAGE (D). (A) HUVECs were incubated with 0.1% DMSO in the presence or absence of 10 μM carboxy-H₂DFFDA for 1 hr, and then treated with or without the indicated concentrations of GLAP in the presence or absence of 5 μg/ml RAGE-Ab. After 10 min, intracellular superoxide generation was measured. * and **, p < 0.05 and p < 0.01 compared to the value without treatment, respectively. #, p < 0.05 compared to the value with 10 μg/ml GLAP. N = 4 per group. (B) HUVECs were treated with or without 10 μg/ml GLAP in the presence or absence of 5 μg/ml RAGE-Ab for 4 hr. Then total RNAs were extracted, transcribed and amplified by real-time PCR. Data were normalized by the intensity of 18S mRNA-derived signals and then related to the value without treatment. #, p < 0.05 compared to the value with 10 μg/ml GLAP. N = 3-6 per group. (C) Secondary structure of GLAP-aptamer. Phoshorothioate linkages are shown as bold line. (D) GLAP at 1, 2 and 3 μg/ml was added on vRAGE-immobilized reaction vessel. The time course of the frequency decrease of bound vRAGE on the QCM was monitored. Two-independent experiments were performed.