Fig. 2.

Effect of AGE-BSA administration on p-ERK1/2 and p-JNK levels. a Western immunoblotting and corresponding densitometric analysis showing abundance of p-ERK1/2 and p-JNK in human aortic endothelial cells (HAECs) after treatment with increasing concentrations of AGE-BSA (100, 200 μg/mL) at various time points (1–72 h). Control, untreated HAECs cultivated for the entire 72-h time course. b Western immunoblotting and corresponding densitometric analysis showing specificity of AGE-BSA-mediated p-ERK1/2 and p-JNK activation (200 μg/mL, 72 h) after pre-treatment with p-ERK1/2 and p-JNK inhibitors (PD98059—50 μM and SP600125—20 μM, respectively). Control, HAECs pre-incubated with solvent alone (DMSO) prior to AGE-BSA treatment (200 μg/mL, 72 h). c Western immunoblotting and corresponding densitometric analysis showing reduction in p-ERK1/2 and p-JNK abundance after pre-incubation of cells with anti-RAGE blocking (neutralizing) antibody (dilution 1:100) for 90 min prior to treatment with AGE-BSA (200 μg/mL) for 72 h. Control, HAECs pre-incubated with goat anti-serum prior to AGE-BSA treatment (200 μg/mL, 72 h). d Detection of RAGE protein expression in HAECs by FACS analysis before and after AGE-BSA administration (200 μg/mL, 72 h). Representative results and corresponding quantification data of one experiment are shown