Th2-predominant inflammation and blockade of IFN-γ signaling induce aneurysms in allografted aortas
J. Clin. Invest. Koichi Shimizu, et al. 114:300 doi:10.1172/JCI19855 [Go to this article.]

Figure 5
Western blot for MMP-12, elastase colorimetric assay, and gelatin and casein zymography. (A) Representative gel image of Western blot analysis. Protein extracts (20 μg/lane) of the aortic grafts of WT (n = 6), GRKO (n = 6), or DKO (n = 6) recipients analyzed by Western blot for MMP-12. The gel images represent qualitatively similar results. (B) The elastase colorimetric assay shows significantly greater elastase activity in the proteins extracted from allografts in GRKO hosts (n = 6) compared with proteins extracted from allografts in WT (n = 6) or DKO (n = 6) hosts. After anti–MMP-12 immunoprecipitation (IP) (n = 6), the proteins from allografts from GRKO recipients had significantly reduced elastase activity, which indicates that the majority of elastolytic activity in those allografts derives from MMP-12. Bar shows mean ± SEM; *P < 0.0001. (C and D) Representative gel images of gelatin zymogram (C) and casein zymogram (D). Protein extracts (20 μg/lane) of the aortic grafts of WT (n = 6), GRKO (n = 6), or DKO (n = 6) recipients analyzed by (C) gelatin- or (D) casein-zymogram for MMPs. The gel images represent qualitatively similar results. We could detect only 92 kDa and 72 kDa active bands from GRKO recipient allografts in the gelatin zymogram (C) and only 20 kDa active band from GRKO recipient allografts in the casein zymogram (D).