Changes in mouse aortic VCAM1 protein levels with age and modulation of
Vcam1
gene expression by mtROS levels. (A) Representative mouse aortic sections immunostained for VCAM1. Scale is 100 μm. (B) Immunohistochemistry scoring of VCAM1 levels (mean±SEM, n=7). (C) Real-time PCR analysis of Vcam1 mRNA expression in VSMCs of young and aged wild-type and p47phox−/− mice treated with 1 U/ml thrombin, 10 μM MitoTEMPO, and thrombin in the presence of MitoTEMPO for 4 h. Data, normalized to 18S RNA expression and relative to the untreated wild-type 4 mo VSMCs, are the mean±SEM of four independent experiments. (D) Representative images of immunofluorescent staining for VCAM1 in VSMCs of young and aged wild-type and p47phox−/− mice treated with 1 U/ml thrombin, 10 μM MitoTEMPO, and thrombin in the presence of MitoTEMPO for 16 h. MitoTEMPO, (2-(2,2,6,6-Tetramethylpiperidin-1-oxyl-4-ylamino)-2-oxoethyl)triphenylphosphonium chloride; NS, not significant; VCAM, vascular cell adhesion molecule.