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Richard R. Keen, MD, Kevin D

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1 Interleukin-1ß induces differential gene expression in aortic smooth muscle cells* 
Richard R. Keen, MD, Kevin D. Nolan, MD, MPH, Maria Cipollone, BS, Elizabeth Scott, MD, Vera P. Shively, MS, James S.T. Yao, MD, PhD, William H. Pearce, MD  Journal of Vascular Surgery  Volume 20, Issue 5, Pages (November 1994) DOI: /S (94) Copyright © 1994 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

2 Fig. 1 Collagenase (MMP-1) and tissue inhibitor of metalloproteinase-2 (TIMP-2) gene expression in human aneurysmal smooth muscle cells. Total ribonucleic acid from cultured aneurysmal aortic smooth muscle cells treated with serum-free conditions for 24 hours followed by 24-hour treatment with either no added interleukin-1ß (IL-1ß) (aneurysmal aortic smooth muscle cell control) or human interleukin-1ß at 100 pg/ml or 1000 pg/ml. These early passage (P4) smooth muscle cells were derived from abdominal aortic aneurysm of 76-year-old man. Experiments were performed in triplicate. Ribonucleic acid was run on formaldehyde agarose gel, blotted on nylon membrane, and hybridized simultaneously with 32P-labeled human cDNA probes for collagenase and tissue inhibitor of metalloproteinase-2. 28S and 18S denote position of ribosomal ribonucleic acid. Journal of Vascular Surgery  , DOI: ( /S (94) ) Copyright © 1994 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

3 Fig. 2 Collagenase (MMP-1) gene expression in human normal aorta smooth muscle cells (NA-SMC). Total ribonucleic acid from cultured NA-SMC treated with serum-free conditions to 24 hours followed by 24-hour treatment with either no added interleukin-1ß (NA-SMC control) or human interleukin-1ß at 100 pg/ml or 1000 pg/ml. These early passage (P3) SMCs were derived from normal abdominal aorta of 53-year-old woman. Experiments were performed in triplicate. Ribonucleic acid was isolated and analyzed by Northern hybridization as in Fig. 1, with human cDNA for collagenase. Journal of Vascular Surgery  , DOI: ( /S (94) ) Copyright © 1994 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

4 Fig. 3 Collagenase (MMP-1) gene expression in human aneurysmal aorta smooth muscle cells (AA-SMC). Total ribonucleic acid from cultured AA-SMC treated with serum-free condition for 24 hours followed by 24 hour treatment with either no added interleukin-1ß (AA-SMC control) or human IL-1ß at 10 pg/ml. These early passage (P4) SMCs were derived from abdominal aortic aneurysm of 77-year-old woman. Ribonucleic acid was isolated and analyzed by Northern hybridization as in Fig. 1 with human cDNA probe for collagenase. Journal of Vascular Surgery  , DOI: ( /S (94) ) Copyright © 1994 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

5 Fig. 4 Type I procollagen (COL 1) gene expression in human aneurysmal aorta smooth muscle cells (AA-SMC). Total ribonucleic acid from cultured AA-SMC derived from 76-year-old man were treated for 24 hours with either no added interleukin-1ß (AA-SMC control); interleukin-1ß at 100 pg/ml; interleukin-1ß at 1000 pg/ml; or interleukin-1ß at 10 ng/ml. Northern hybridization was performed with a labeled cDNA probe for human type I procollagen. Journal of Vascular Surgery  , DOI: ( /S (94) ) Copyright © 1994 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

6 Fig. 5 Type I procollagen (COL 1) gene expression in human normal aortic smooth muscle cells (NA-SMC). Total ribonucleic acid from cultured NA-SMC derived from 59-year-old woman was treated for 24 hours with either no added interleukin-1ß (NA-SMC control); interleukin-1ß at 10 pg/ml; interleukin-1ß at 100 pg/ml; or interleukin-1ß at 1000 pg/ml. Northern hybridization was performed with labeled human cDNA probe for type I procollagen. Journal of Vascular Surgery  , DOI: ( /S (94) ) Copyright © 1994 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

7 Fig. 6 Tissue inhibitor of metalloproteinase-1 (TIMP-1) gene expression in human aneurysmal aorta smooth muscle cells (AA-SMC). Total ribonucleic acid from cultured AA-SMC from 77-year-old woman was treated for 24 hours with either no added interleukin-1ß (AA-SMC control); interleukin-1ß at 10 pg/ml; interleukin-1ß at 100 pg/ml; or interleukin-1ß at 1000 pg/ml. Ribonucleic acid was analyzed by Northern hybridization with human cDNA probe for TIMP-1. Journal of Vascular Surgery  , DOI: ( /S (94) ) Copyright © 1994 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

8 Fig. 7 Tissue inhibitor of metalloproteinase-1 (TIMP-1) gene expression in normal aorta smooth muscle cells (NA-SMC). Total ribonucleic acid from cultured NA-SMC derived from 53-year-old woman was treated for 24 hours with either no added interleukin-1ß (NA-SMC control); interleukin-1ß at 100 pg/ml; or interleukin-1ß at 1000 pg/ml. Northern hybridization was performed with labeled cDNA probe for TIMP-1. Journal of Vascular Surgery  , DOI: ( /S (94) ) Copyright © 1994 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

9 Fig. 8 Effects of increased concentrations of interleukin-1ß on percent change in steady-state messenger ribonucleic acid levels as compared with (unstimulated) control cells for collagenase (MMP-1), tissue inhibitor of metalloproteinase-1 (TIMP-1), and type I procollagen (COL 1) in smooth muscle cells explanted and cultured from human aneurysmal aorta (AA-SMC). Values are mean ± SE. Journal of Vascular Surgery  , DOI: ( /S (94) ) Copyright © 1994 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

10 Fig. 9 Effects of increasing concentrations of interleukin-1ß on percent change in steady-state messenger ribonucleic acid levels compared with (unstimulated) control cells for collagenase (MMP-1), tissue inhibitor of metalloproteinase-1 (TIMP-1), and type I procollagen (COL-1) in human normal aorta smooth muscle cells (NA-SMC) explanted and cultured from elderly organ donors. Values are mean ± SE. Journal of Vascular Surgery  , DOI: ( /S (94) ) Copyright © 1994 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

11 Fig. 10 72kDa Type IV collagenase (MMP-2) gene expression in human aorta smooth muscle cells (AA-SMC). Total ribonucleic acid from cultured AA-SMC treated with serum-free conditions for 24 hours followed by 24-hour treatment with either no added human platelet-derived growth factor (PDGF); PDGF at 100 pg/ml; or PDGF at 1000 pg/ml. These early passage (P2) cells were derived from abdominal aortic aneurysm of 77-year-old woman. Northern hybridization was performed with labeled cDNA probe for 72 kDa type IV collagenase (MMP-2). Journal of Vascular Surgery  , DOI: ( /S (94) ) Copyright © 1994 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

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