Characterization of primary and restenotic atherosclerotic plaque from the superficial femoral artery: Potential role of Smad3 in regulation of SMC proliferation

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Characterization of primary and restenotic atherosclerotic plaque from the superficial femoral artery: Potential role of Smad3 in regulation of SMC proliferation Rachel S. Edlin, BS, Shirling Tsai, MD, Dai Yamanouchi, MD, PhD, Chunjie Wang, MD, Bo Liu, PhD, K. Craig Kent, MD Journal of Vascular Surgery Volume 49, Issue 5, Pages 1289-1295 (May 2009) DOI: 10.1016/j.jvs.2008.11.096 Copyright © 2009 Society for Vascular Surgery Terms and Conditions

Fig 1 Immunohistochemical analysis of human atherosclerotic and restenotic lesions. Immunohistochemistry of primary atherosclerotic (n = 5) and restenotic lesions (n = 3). Representative sections are shown. Staining for (A) α-actin, (B) PCNA, (C) CD68, and (D) CD45. Magnification ×400. E, Quantification of positive immunostaining in restenotic or primary atherosclerotic lesions expressed as a ratio of positive/total cells ± standard error (*P < .001 restenotic compared with primary lesions). Journal of Vascular Surgery 2009 49, 1289-1295DOI: (10.1016/j.jvs.2008.11.096) Copyright © 2009 Society for Vascular Surgery Terms and Conditions

Fig 2 Smad3 staining of human atherosclerotic and restenotic lesions. Smad3 staining of primary atherosclerotic lesions (n = 5) and restenotic lesions (n = 3). Representative sections are shown. Magnification ×400. Journal of Vascular Surgery 2009 49, 1289-1295DOI: (10.1016/j.jvs.2008.11.096) Copyright © 2009 Society for Vascular Surgery Terms and Conditions

Fig 3 Colocalization of Smad3 with α-actin and PCNA. Double immunofluorescent staining of restenotic plaque. A, Staining reveals (a) nuclei (blue), (b) nuclear Smad3 (green), (c) α -actin (red), and (d) colocalization (nuclear green surrounded by cytoplasmic red) (white arrows). B, Staining reveals (a) nuclei (blue), (b) nuclear Smad3 (green), (c) nuclear PCNA (red), and (d) colocalization (yellow) (white arrows). Journal of Vascular Surgery 2009 49, 1289-1295DOI: (10.1016/j.jvs.2008.11.096) Copyright © 2009 Society for Vascular Surgery Terms and Conditions

Fig 4 The effect of Smad3 on human SMC proliferation. A, Western blotting of human aortic SMCs demonstrating the effects of Smad3 overexpression via AdSmad3 infection and Smad3 downregulation via transient transfection with Smad3 siRNA. B, 3H-thymidine incorporation in human aortic SMCs following overexpression or inhibition of Smad3 (n = 3, *P < .001 compared with AdLacZ or scrambled control). Journal of Vascular Surgery 2009 49, 1289-1295DOI: (10.1016/j.jvs.2008.11.096) Copyright © 2009 Society for Vascular Surgery Terms and Conditions

Fig 5 The effect of Smad3 on SMC apoptosis. Vascular SMCs were infected with AdLacZ or AdSmad3, then lysed and analyses for apoptosis using ELISA. *P < .05 compared with negative control. Journal of Vascular Surgery 2009 49, 1289-1295DOI: (10.1016/j.jvs.2008.11.096) Copyright © 2009 Society for Vascular Surgery Terms and Conditions