Effects of AGEs and mechanical stretching on phosphorylation of ERK1/2 in VSMCs.

<p>80% confluent VSMCs were serum-starved for 48 hours and treated with AGEs and/or mechanical stretching as indicated for 10 minutes. They were then harvested for Western blot analysis. BSA was used as a negative control. Panels A and B show that AGEs induced ERK activation in VSMCs in a dose- and time-dependent manner. Panel C shows that mechanical stretching induced ERK activation in VSMCs in an elongation-dependent manner. Panel D shows that combined stimulation synergistically activated ERKs in VSMCs. Beta-actin served as an internal control. Graphs E, F, G, and H show the statistical results of phosphorylated-ERK (pERK) levels (ERK activity) corresponding to Panels A, B, C, and D from three independent experiments. Both (Panel I) incubation with AGEs or (Panel K) stimulation by mechanical stretching were found to elevate the expression of RAGE in a time-dependent manner. *P<0.05 <i>versus</i> negative control (NC), #P<0.05 <i>versus</i> AGE or stretch stress (SS).