Article Title: β-Cyclodextrin Inhibits Monocytic Adhesion to Endothelial Cells through Nitric Oxide-Mediated Depletion of Cell Adhesion Molecules
Figure Lengend Snippet: Protein kinase Cε mediates phosphorylation of endothelial nitric oxide synthetase at threonine-495 in bovine aortic endothelial cells. ( A ). Bovine aortic endothelial cells (BAECs) were incubated with 400 μg/mL β-cyclodextrin (β-CD) for the time-course experiment. Then the cell lysates were immunoblotted with anti-caveolin and p-caveolin antibodies. The quantified data are shown in the bottom panel (mean ± SE, n = 3). ( B ). BAECs were pretreated with 400 μg/mL β-CD for the indicated times and then incubated with 1 μM A23187 for 20 min. Subsequently, intracellular [Ca 2+ ] was measured by fluorescence spectroscopy (Ex: 490 nm, Em: 525 nm) 30 min after incubation with 5 μM Fluo-8AM (mean ± SE, n = 3). ( C ). BAECs were incubated with 100 nM phorbol-12-myristate-13-acetate (PMA) in the time-course experiment. Cell lysates were immunoblotted with anti-eNOS and various anti-p-T495 eNOS antibodies. The data were quantified using densitometry (bottom panel, mean ± SE, n = 3). * p
Article Snippet: The activators used were phorbol 12-myristate 13-acetate (PMA, 100 nmol/L, Enzo Life Sciences Inc., Farmingdale, NY, USA) and glycerol-1,3-dipalmitate (diacylglycerol (DAG), 100 μmol/L, Sigma-Aldrich, St. Louis, MO, USA).
Techniques: Incubation, Fluorescence, Spectroscopy