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Figure 6: Analysis of the expression of the AGE/RAGE axis in HUVECs. The HUVECs were treated with 400 μM MG and glycine (0.5 mM-4 mM) for 72 h. (a) and (b) show the immunofluorescence analysis of AGE expression levels in HUVECs, magnification 400x. The scale bar indicates 20 μm. The images are the representative of three independent experiments. (c) Intracellular AGE levels determined by ELISA. The experiment was performed in triplicate and repeated three times. The expression levels of RAGE (d), Nox4 (e), and NF-κB p65 (f) in HUVECs were determined by western blot. The experiment was performed in four independent experiments. (g) and (h) show the estimation of intracellular ROS generation by flow cytometry. The images are the representative of four independent experiments. Control: cells treated with 5.5 mM glucose. MG: methylglyoxal. Gly: glycine. , compared with the control group. , , compared with cells treated with 400 μM MG.