Figure 3

Secondary structure of AGE-aptamer (A), binding affinities of GLAP to AGE-aptamer, GLAP-aptamer, or vRAGE (B-E) and effects of AGE- or GLAP-aptamer on inflammatory and thrombogenic gene expression in GLAP-exposed HUVECs (F-K). (A) Secondary structure of AGE-aptamer. Phoshorothioate linkages are shown as bold line. AGE-aptamer (B), GLAP ((C) and (D)) or recombinant vRAGE (E) was immobilized on a QCM surface. After adding GLAP ((B) and (E)), GLAP-aptamer (C) or each aptamer (D) to reaction vessel, the time course of the frequency decrease of bound AGE-aptamer (B), bound GLAP ((C) and (D)), or bound vRAGE (E on the QCM was monitored. Under no serum conditions, solid line and circule. Under 2% fetal bovine serum conditions, dashed line and triangle. N = 3 per group. (F)-(K) HUVECs were treated with or without 10 μg/ml GLAP in the presence or absence of 100 nM each aptamer 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 obtained with control. # and ##, p < 0.05 and p < 0.01 compared to the value with 10 μg/ml GLAP alone, respectively. N = 4 per group.