Journal: Circulation

Article Title: Angiopoietin-1 protects against the development of cardiac allograft arteriosclerosis.

doi: 10.1161/01.cir.0000054623.35669.3f

Figure Lengend Snippet: Figure 1. Kinetics of Ang1 (A), Ang2 (B), and Tie2 (C) immunoreactivity in acute and chronic rejection of rat cardiac allografts. Ang1 is given as number of positive cells per cross section, whereas Ang2 and Tie2 are given as number of positive capillaries and postcapillary venules per cross section. Data are meanSEM, by Student t test. N values are the following: nor- mal heart, 5; acute syngraft, 5; acute allograft, 5; chronic syngraft, 6; and chronic allograft, 11.

Article Snippet: Cryostat sections were stained using a peroxidase ABC method (Vectastain Elite ABC Kit, Vector Laboratories), and the reaction was revealed by 3-amino-9-ethylcarbazole (Vectastain).19 Antibodies and dilutions used were as follows: Ang1 (10 g/mL, RDIANGIOPO1Xabr) from Research Diagnostics; Ang2 (2 g/mL, sc-7017), Tie2 (2 g/mL, sc-9026), and CD34 (2 g/mL, sc-7324) from Santa Cruz Biotechnology; CD4 (5 g/mL, 22021D), CD8 (5 g/mL, 22071D), ED1 (5 g/mL, 22451D), and interleukin (IL)– 2R (5 g/mL, 22090D) from PharMingen; RECA-1 (1:10, MCA970) from Serotec; leukocyte common antigen (LCA, 1:100, clone OX10) and major histocompatibility complex (MHC) class II (1:100, clone OX6) from Sera Laboratory; vascular cell adhesion molecule (VCAM)–1 (10 g/mL, MMS-141P) from BAbCO (CRP, Inc); and ICAM-1 (10 g/mL, clone 1A29) from Seikagaku.

Techniques:

Journal: Circulation

Article Title: Angiopoietin-1 protects against the development of cardiac allograft arteriosclerosis.

doi: 10.1161/01.cir.0000054623.35669.3f

Figure Lengend Snippet: Figure 2. Localization of Ang1, Ang2, and Tie2 immu- noreactivity in chronic rejection of rat cardiac allografts. In normal hearts (A and B), only a few Ang1- immunoreactive cells were detected in cardiac mesen- chyme (A), whereas mild Ang2 immunoreactivity was detected in capillaries and postcapillary venules (B). In chronic allografts (C through H), Ang2 immunoreactivity was upregulated in capillaries and postcapillary venules (C; inset shows a longitudinally cut capillary, and arrows point to perivascular infiltrated mononuclear cells). Ang2 was also expressed in some cardiomyocytes (D) and infiltrating mononuclear cells (E) in chronic allografts. Arterial endothelium did not express Ang2 (E, arrow), but Ang2 immunoreactivity was detected in intimal neo- capillaries of some totally occluded arteries (F), which also stained for the endothelial cell marker RECA-1 (F, inset) and Tie2 (G). Tie2 immunoreactivity was detected mainly in capillaries and postcapillary venules (H). Rat uterus was used as a positive control and glandular epithelium showed intense immunoreactivity for Ang2 (I), whereas incubation of the primary antibody with an excess of Ang2 peptide resulted in a lack of any spe- cific immunoreactivity (I, inset). Sections were counter- stained with Mayer’s hemalum. Dotted line indicates internal elastic lamina. Scale bars20 m.

Article Snippet: Cryostat sections were stained using a peroxidase ABC method (Vectastain Elite ABC Kit, Vector Laboratories), and the reaction was revealed by 3-amino-9-ethylcarbazole (Vectastain).19 Antibodies and dilutions used were as follows: Ang1 (10 g/mL, RDIANGIOPO1Xabr) from Research Diagnostics; Ang2 (2 g/mL, sc-7017), Tie2 (2 g/mL, sc-9026), and CD34 (2 g/mL, sc-7324) from Santa Cruz Biotechnology; CD4 (5 g/mL, 22021D), CD8 (5 g/mL, 22071D), ED1 (5 g/mL, 22451D), and interleukin (IL)– 2R (5 g/mL, 22090D) from PharMingen; RECA-1 (1:10, MCA970) from Serotec; leukocyte common antigen (LCA, 1:100, clone OX10) and major histocompatibility complex (MHC) class II (1:100, clone OX6) from Sera Laboratory; vascular cell adhesion molecule (VCAM)–1 (10 g/mL, MMS-141P) from BAbCO (CRP, Inc); and ICAM-1 (10 g/mL, clone 1A29) from Seikagaku.

Techniques: Staining, Marker, Positive Control, Incubation

Journal: Circulation

Article Title: Angiopoietin-1 protects against the development of cardiac allograft arteriosclerosis.

doi: 10.1161/01.cir.0000054623.35669.3f

Figure Lengend Snippet: Figure 3. Functionality of Ad.Ang1 in vitro. Metabolically labeled cells were transfected with Ad.lacZ or Ad.Ang1, and culture medium was subjected to immunoprecipitation with Myc- specific antibody or soluble Tie2-immunoglobulin fusion protein. Gel electrophoresis concluded that Ad.Ang1-transfected cells produce recombinant Ang1 (A), which also binds to Tie2 recep- tor (B).

Article Snippet: Cryostat sections were stained using a peroxidase ABC method (Vectastain Elite ABC Kit, Vector Laboratories), and the reaction was revealed by 3-amino-9-ethylcarbazole (Vectastain).19 Antibodies and dilutions used were as follows: Ang1 (10 g/mL, RDIANGIOPO1Xabr) from Research Diagnostics; Ang2 (2 g/mL, sc-7017), Tie2 (2 g/mL, sc-9026), and CD34 (2 g/mL, sc-7324) from Santa Cruz Biotechnology; CD4 (5 g/mL, 22021D), CD8 (5 g/mL, 22071D), ED1 (5 g/mL, 22451D), and interleukin (IL)– 2R (5 g/mL, 22090D) from PharMingen; RECA-1 (1:10, MCA970) from Serotec; leukocyte common antigen (LCA, 1:100, clone OX10) and major histocompatibility complex (MHC) class II (1:100, clone OX6) from Sera Laboratory; vascular cell adhesion molecule (VCAM)–1 (10 g/mL, MMS-141P) from BAbCO (CRP, Inc); and ICAM-1 (10 g/mL, clone 1A29) from Seikagaku.

Techniques: In Vitro, Metabolic Labelling, Labeling, Transfection, Immunoprecipitation, Nucleic Acid Electrophoresis, Recombinant