Journal:

Article Title: A vascular bed-specific pathway regulates cardiac expression of endothelial nitric oxide synthase

doi:

Figure Lengend Snippet: Expression of the 1600eNOS promoter is induced by myocyte-derived signaling pathways in vitro. MEEP cells were stably transfected with 1600eNOS LUC, 744eNOS LUC, and 166 eNOS LUC as described in Methods. Cells were starved in 0.2% FCS for 24 h and then incubated for 4 h in conditioned medium from HEK, HepG2, cardiac myocytes (MYO), skeletal myoctes (SKM), or brain astrocytes (AST) in the absence (a) or presence (b) of anti-PDGF antibodies, or in media supplemented with PDGF, VEGF, bFGF, and EGF at concentrations of 10 ng/ml (b). The results show the mean and SD of luciferase light units obtained in triplicate from one representative experiment. The 1600eNOS LUC construct was induced 4.8-fold by cardiac myocyte–conditioned medium, 4.3-fold by skeletal muscle–conditioned medium, 3-fold by astrocyte-conditioned medium, 2.4-fold by PDGF-AB, and 3-fold by cardiac myocyte–conditioned medium containing anti-PDGF antibodies (b). The 744eNOS LUC construct was induced 2.2-fold, and the 166eNOS LUC construct 2.7-fold, by myocyte-conditioned medium (c and d, respectively). *P < 0.05 vs. basal activity; #P < 0.05 vs. cardiac myocyte–conditioned medium. bFGF, basic fibroblast growth factor; EGF, epidermal growth factor; MEEP, mouse embryonic endothelial progenitor; PDGF, platelet-derived growth factor; VEGF, vascular endothelial growth factor.

Article Snippet: Alternatively, the cells were incubated for 4–24 h with conditioned medium from cardiac myocytes, skeletal myocytes, brain astrocytes, or HEK or HepG2 cells (70% confluence and medium 2–3 days old) in the absence or presence of neutralizing antibodies to PDGF, VEGF, and bFGF (R&D Systems).

Techniques: Expressing, Derivative Assay, In Vitro, Stable Transfection, Transfection, Incubation, Luciferase, Construct, Activity Assay

Journal:

Article Title: A vascular bed-specific pathway regulates cardiac expression of endothelial nitric oxide synthase

doi:

Figure Lengend Snippet: eNOS RNA levels are increased by PDGF. BAECs were starved in 0.5% FCS for 24 h and then incubated with either VEGF, EGF, bFGF, or PDGF-AB at 10 ng/ml for 4 h. Total RNA (5 μg) was subjected to agarose electrophoresis. (a) Autoradiogram with an eNOS cDNA probe. (b) The ethidium-stained 28S ribosomal band. BAECs, bovine aortic endothelial cells.

Article Snippet: Alternatively, the cells were incubated for 4–24 h with conditioned medium from cardiac myocytes, skeletal myocytes, brain astrocytes, or HEK or HepG2 cells (70% confluence and medium 2–3 days old) in the absence or presence of neutralizing antibodies to PDGF, VEGF, and bFGF (R&D Systems).

Techniques: Incubation, Electrophoresis, Staining