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Recombinant human CD253 used to assess induction of MMP-9 and Timp-1 by human trail on murine aortic smooth muscle cells in vitro
Induction of Mmp-9 and Timp-1 mRNA by Trail via the JNK or NF-κB Signaling Pathway. (a, b, d, e) Relative expression levels of Mmp-9 (a, b) and Timp-1 (d, e) mRNA in mouse aortic SMCs within a culture system induced by rh-TRAIL (10 ng/ml). SP-600125 (3–30 μM) (a) and AS-601245 (0.2–20 μM) (b) were used to inhibit the JNK signaling pathway. MG-132 (0.2–20 μM) (d) and BAY11-7082 (1–10 μM) (e) were used to inhibit the NF-κB pathway. Data are presented as mean ± SD; *p<0.05 or **p<0.01, as compared to controls or TRAIL alone. Samples were in triplicate for each group. (c, f) Representative double immunofluorescent staining images of abdominal aortas in wild-type and Opg-KO mice at six weeks, with pJNK (panels A-b, B-b, and C-b in c) or pNF-κB (panels D-b, E-b, and F-b in f) in green, and Mmp-9 (panels A-c, B-c, and C-c in c) or Timp-1 (panels D-c, E-c, and F-c in f) in red. pJNK signal merged with Mmp-9 (panels A-d, B-d, and C-d in c). pNF-κB signal merged with Timp-1 (panels A-d, B-d, and C-d in f). Rectangles in panels B and E show magnified areas in C and F, respectively. Nuclei are stained with DAPI in blue (m: medial layer; a: adventitia). Scale bars in panels A, B, D, and E represent 100 μm. Scale bars in panels C and F represent 5μµm.
From: Bumdelger B, Kokubo H, Kamata R, Fujii M, Yoshimura K, Aoki H, et al. (2016) Osteoprotegerin Prevents Development of Abdominal Aortic Aneurysms. PLoS ONE 11(1): e0147088.