Journal: Hypertension

Article Title: Calpain-1 Regulation of Matrix Metalloproteinase 2 Activity in Vascular Smooth Muscle Cells Facilitates Age-Associated Aortic Wall Calcification and Fibrosis

doi: 10.1161/hypertensionaha.112.196840

Figure Lengend Snippet: Figure 3. Overexpression of calpain-1 induces collagen type I, II, and III and reduces elastin expression levels in vascular smooth muscle cells (VSMC). A, Representative Western blots of collagen I and collagen III in VSMC after infection by GFP or CANP1 adenoviruses for 48 hours. B, Photomicrographs (×400) of collagen II protein staining (brown color) from young (8-month) and old (30-month) rat aortic walls. Nuclei are counterstained with hematoxylin (blue color). L indicates lumen; M, media. C and D, Representative Western blots and average protein levels of collagen II (C) and elastin (D) in young VSMC infected with GFP or CANP1 for 48 hours. *P<0.05 compared with GFP-infected 8-month VSMC, n=3.

Article Snippet: Primary antibodies used for immunostaining (IS) and Western blotting (WB) Antibody Species Titer for IS Titer for WB Source -actin Mouse 1:10000 Sigma-Aldrich Co. LLC, St. Louis, MO calpain-1 Rabbit 1:100 1:2500 Sigma-Aldrich Co. LLC, St. Louis, MO collagen I Rabbit 1:1000 Rockland Antibodies & Assays, Gilbertsville, PA collagen II Rabbit 1:50 1:1000 Abcam, Cambridge, MA collagen III Rabbit 1:1000 Rockland Antibodies & Assays, Gilbertsville, PA elastin Rabbit 1:1000 Millipore, Billerica, MA MMP2 Mouse 1:50 1:1000 Millipore, Billerica, MA MT1MMP Rabbit 1:1000 Millipore, Billerica, MA osteonectin Rabbit 1:1000 Millipore, Billerica, MA osteoponin Rabbit 1:1000 Millipore, Billerica, MA Smad 2/3 Mouse 1:1000 Cell Signaling Technology, Inc, Danvers, MA TGF-1 Rabbit 1:500 Santa Cruz Biotechnology, Inc, Santa Cruz, CA TIMP2 Goat 1:500 R&D Systems, Inc. Minneapolis, MN 9 Figure S1: Interactions between calpain-1 and MMP2.

Techniques: Over Expression, Expressing, Western Blot, Infection, Staining

Journal: Hypertension

Article Title: Calpain-1 Regulation of Matrix Metalloproteinase 2 Activity in Vascular Smooth Muscle Cells Facilitates Age-Associated Aortic Wall Calcification and Fibrosis

doi: 10.1161/hypertensionaha.112.196840

Figure Lengend Snippet: Figure 6. Overexpression of calpain-1 regulates extracellular matrix remodeling, particularly fibrosis and vascular calcification in cultured carotid artery rings. Representative Western blot of tissue inhibitor of metalloprotease 2 (TIMP2), calpain-1, elastin, transforming growth factor-β1 (TGFβ1), (A) Smad 2/3, collagen I, II, III, (B) and osteopontin (OPN) and osteonectin (ON) (C) in cultured carotid artery rings isolated from 8-month-old rat after infection by GFP or CANP1 or TIMP2 adenoviruses for 48 hours.

Article Snippet: Primary antibodies used for immunostaining (IS) and Western blotting (WB) Antibody Species Titer for IS Titer for WB Source -actin Mouse 1:10000 Sigma-Aldrich Co. LLC, St. Louis, MO calpain-1 Rabbit 1:100 1:2500 Sigma-Aldrich Co. LLC, St. Louis, MO collagen I Rabbit 1:1000 Rockland Antibodies & Assays, Gilbertsville, PA collagen II Rabbit 1:50 1:1000 Abcam, Cambridge, MA collagen III Rabbit 1:1000 Rockland Antibodies & Assays, Gilbertsville, PA elastin Rabbit 1:1000 Millipore, Billerica, MA MMP2 Mouse 1:50 1:1000 Millipore, Billerica, MA MT1MMP Rabbit 1:1000 Millipore, Billerica, MA osteonectin Rabbit 1:1000 Millipore, Billerica, MA osteoponin Rabbit 1:1000 Millipore, Billerica, MA Smad 2/3 Mouse 1:1000 Cell Signaling Technology, Inc, Danvers, MA TGF-1 Rabbit 1:500 Santa Cruz Biotechnology, Inc, Santa Cruz, CA TIMP2 Goat 1:500 R&D Systems, Inc. Minneapolis, MN 9 Figure S1: Interactions between calpain-1 and MMP2.

Techniques: Over Expression, Cell Culture, Western Blot, Isolation, Infection