Comparison of cell-type-specific vs transmural aortic gene expression in experimental aneurysms Eiketsu Sho, MD, PhD, Mien Sho, MD, Hiroshi Nanjo, MD,

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Comparison of cell-type-specific vs transmural aortic gene expression in experimental aneurysms Eiketsu Sho, MD, PhD, Mien Sho, MD, Hiroshi Nanjo, MD, PhD, Koichi Kawamura, PhD, Hirotake Masuda, MD, PhD, Ronald L. Dalman, MD Journal of Vascular Surgery Volume 41, Issue 5, Pages 844-852 (May 2005) DOI: 10.1016/j.jvs.2005.02.027 Copyright © 2005 The Society for Vascular Surgery. Terms and Conditions

Fig 1 Representative experimental aneurysm samples. A, Hematoxylin and eosin-stained AAA tissue before laser capture microdissection; B, AAA after laser capture microdissection; C, immunohistochemical staining for smooth muscle cells (SM-α actin); D, immunohistochemical staining for macrophages (ED-1) (original magnification: A and B, ×40; C and D, ×200); E, α-actin PCR product bands (101 base pair) were detected in 2% agarose gel after 25 cycles of PCR from aRNA showing all samples containing constant aRNA. C, control; M, mark. Journal of Vascular Surgery 2005 41, 844-852DOI: (10.1016/j.jvs.2005.02.027) Copyright © 2005 The Society for Vascular Surgery. Terms and Conditions

Fig 2 Ultrastructural analysis of experimental AAA. Endothelial cell denudation and luminal microthrombi present day 1 after PPE infusion (A and B). Apoptotic medial smooth muscle cells (C and D) were conspicuous for distinct nuclear chromatin condensation (arrows) and vacuolar cytoplasm (arrowheads). SM, smooth muscle cell. Journal of Vascular Surgery 2005 41, 844-852DOI: (10.1016/j.jvs.2005.02.027) Copyright © 2005 The Society for Vascular Surgery. Terms and Conditions

Fig 3 Flow-dependant cell proliferation and SMC apoptosis. All panels represent specific cellular indices as functions of time and flow conditions (LF-, NF-, or HF-AAA). A, BrdU-labeled endothelial cells/cross-sectional lumen (CSL); B, BrdU-labeled SMCs/cross-sectional area (CSA); C, Apoptotic SMCs/CSA; D, Number of ECs/CSL; E, Number of SMCs/high-power field; F, Number of macrophages/CSA. *P < .05 vs NF- and LF-AAA; †P < .05 vs LF-AAA. Journal of Vascular Surgery 2005 41, 844-852DOI: (10.1016/j.jvs.2005.02.027) Copyright © 2005 The Society for Vascular Surgery. Terms and Conditions

Fig 4 Flow-dependant cytokine, growth, and transcription factor expression in whole-organ lysates and LCM specimens. Data reported as expression ratios to control aortic tissue. *P < .05 vs NF- and LF-AAA; †P < .05 vs LF-AAA. Journal of Vascular Surgery 2005 41, 844-852DOI: (10.1016/j.jvs.2005.02.027) Copyright © 2005 The Society for Vascular Surgery. Terms and Conditions

Fig 5 Comparison of MMP-2 and MMP-9 gene expression in total AAA tissue and cell-specific areas. Data were reported as the ratio to control aortic tissue. *P < .05 vs NF- and LF-AAA; †P < .05 vs LF-AAA. Journal of Vascular Surgery 2005 41, 844-852DOI: (10.1016/j.jvs.2005.02.027) Copyright © 2005 The Society for Vascular Surgery. Terms and Conditions

Fig 6 MMP-2 and -9 gelatinolytic activity in experimental AAA. C, normal control aorta; pro-form MMPs, band consistent with molecular weight of precursor form of MMP before activation; active-form MMPs, band consistent with molecular weight of MMP after activation. Both pro-form and active-form MMP isozymes have gelatinolytic activity. *P < .01 vs pro-form MMP-2; †P < .01 vs pro-form MMP-9. Journal of Vascular Surgery 2005 41, 844-852DOI: (10.1016/j.jvs.2005.02.027) Copyright © 2005 The Society for Vascular Surgery. Terms and Conditions