Activation of transglutaminase type 2 for aortic wall protection in a rat abdominal aortic aneurysm formation Takashi Munezane, MD, Tomomi Hasegawa,

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Activation of transglutaminase type 2 for aortic wall protection in a rat abdominal aortic aneurysm formation Takashi Munezane, MD, Tomomi Hasegawa, MD, PhD, Suritala, MD, Akiko Tanaka, MD, Kenji Okada, MD, PhD, Yutaka Okita, MD, PhD Journal of Vascular Surgery Volume 52, Issue 4, Pages 967-974 (October 2010) DOI: 10.1016/j.jvs.2010.04.049 Copyright © 2010 Society for Vascular Surgery Terms and Conditions

Fig 1 TG2 expression in AAAs. (A) Relative TG2 mRNA expression to the native aorta. *P < .05, (B) TG2 protein expression and enzyme activity. Bar = 100 μm. AAA, Abdominal aortic aneurysm; AAA prep, AAA-prepared aorta; No prep, non-prepared aorta; TG2, transglutaminase type 2. All data are expressed as mean ± standard error of the mean for six rats per group. Journal of Vascular Surgery 2010 52, 967-974DOI: (10.1016/j.jvs.2010.04.049) Copyright © 2010 Society for Vascular Surgery Terms and Conditions

Fig 2 Messenger RNA expression of proinflammatory cytokines and MMPs in AAAs at 1 and 8 weeks. (A) TNF-α, IL-1β, MMP-2, and MMP-9. (B) TIMP-1 and TIMP-2. AAA, Abdominal aortic aneurysm; AAA prep, AAA-prepared aorta; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; IL, interleukin; MMP, matrix metalloproteinase; No prep, non-prepared aorta; NS, not significant; TIMP, tissue inhibitors of metalloproteinase; TNF, tissue necrosis factor. All data are expressed as mean ± standard error of the mean for six rats per group. Journal of Vascular Surgery 2010 52, 967-974DOI: (10.1016/j.jvs.2010.04.049) Copyright © 2010 Society for Vascular Surgery Terms and Conditions

Fig 3 Double immunofluorescent staining for TG2 (green) and TNF-α (red). Sites of colocalization of TG2 and TNF-α are indicated as a yellow emission. Cell nuclei are counterstained with DAPI (blue). Bar = 100 μm. AAA, Abdominal aortic aneurym; AAA prep, AAA-prepared aorta; DAPI, 4′,6-diamidino-2-phenylindole; No prep, non-prepared aorta; TG2, transglutaminase type 2; TNF, tissue necrosis factor. Journal of Vascular Surgery 2010 52, 967-974DOI: (10.1016/j.jvs.2010.04.049) Copyright © 2010 Society for Vascular Surgery Terms and Conditions

Fig 4 Messenger RNA expression of TNF-α, MMP-2, and MMP-9 after 3-day incubation of isolated AAA segments with exogenous transglutaminase type 2 (TG2) or TG2 inhibitor cystamine (Cys), or without them (Conrol). The AAA segments were harvested at 1 week after AAA preparation in the rat AAA model. Cys, Cysteine; IL, interleukin; MMP, matrix metalloproteinase; NS, not significant; TG2, transglutaminase type 2; TNF, tissue necrosis factor. All data are expressed as mean ± standard error of the mean for six independent analyses. Journal of Vascular Surgery 2010 52, 967-974DOI: (10.1016/j.jvs.2010.04.049) Copyright © 2010 Society for Vascular Surgery Terms and Conditions

Fig 5 Messenger RNA expression of TG2 after 3-day incubation of isolated AAA segments with/without TNF-α. The AAA segments were harvested at 1 week after AAA preparation in the rat AAA model. The data were compared with TG2 mRNA level before tissue culture. TG2, Transglutaminase type 2; TNF, tissue necrosis factor; pre, preculture. All data are expressed as mean ± standard error of the mean for six independent analyses. Journal of Vascular Surgery 2010 52, 967-974DOI: (10.1016/j.jvs.2010.04.049) Copyright © 2010 Society for Vascular Surgery Terms and Conditions