Journal: Physiological Reports

Article Title: Regulation of cell proliferation by the guanosine–adenosine mechanism: role of adenosine receptors

doi: 10.1002/phy2.24

Figure Lengend Snippet: Effects of guanosine on [ 3 H]-thymidine incorporation in human coronary artery vascular smooth muscle cells in the absence and presence of adenosine. Values represent means and SEMs. Two-factor ANOVA indicated a significant interaction between guanosine and adenosine on [ 3 H]-thymidine incorporation ( P < 0.000001). Letter “a” indicates a significant inhibitory response to guanosine at the indicated concentration of adenosine, and “b” indicates that the inhibitory response to guanosine is significantly greater in the presence of adenosine.

Article Snippet: Human coronary artery smooth muscle cells were obtained from Lonza Inc. (Allendale, NJ).

Techniques: Concentration Assay

Journal: Physiological Reports

Article Title: Regulation of cell proliferation by the guanosine–adenosine mechanism: role of adenosine receptors

doi: 10.1002/phy2.24

Figure Lengend Snippet: Effects of adenosine on [ 3 H]-thymidine incorporation in human coronary artery vascular smooth muscle cells in the absence (left panel) and presence (right panel) of guanosine, both without (no DPSPX; top graph) and with (pretreated with DPSPX; bottom graph) 1,3-dipropyl-8-( p -sulfophenyl)xanthine (DPSPX; 100 μmol/L; adenosine receptor antagonist). Values represent means and SEMs. Two-factor ANOVA indicated a significant interaction between guanosine and adenosine on [ 3 H]-thymidine incorporation in the “no DPSPX” group ( P < 0.000001) but not in the “pretreated with DPSPX” group. Letter “a” indicates significantly different from corresponding group without adenosine, and “b” indicates significantly different from corresponding “no guanosine” group.

Article Snippet: Human coronary artery smooth muscle cells were obtained from Lonza Inc. (Allendale, NJ).

Techniques:

Journal: American Journal of Physiology - Cell Physiology

Article Title: Shear stress induces Gα q/11 activation independently of G protein-coupled receptor activation in endothelial cells

doi: 10.1152/ajpcell.00148.2016

Figure Lengend Snippet: Ligand- and shear stress-induced activation of the sphingosine-1-phosphate (S1P3)-Gαq/11 complex. In situ proximity ligation assay (PLA) was performed using antibodies directed against S1P3 and Gαq/11 (A and B), serotonin (5-HT)2A and Gαq/11 (C and D), S1P3 and G protein-coupled receptor kinases (GRK2; E), and S1P3 and β-arrestin-1/2 (F) on human coronary artery endothelial cells (HCAECs) treated with vehicle alone (Control), S1P; 2 μM for 30 s) (A, E, and F), or 5-HT (100 nM for 30 s) (C) and on HCAECs that were either unstimulated (Sham) or subjected to a step change in shear stress (Flow) for the indicated times (B, D, E, and F). Representative confocal images in a single z-plane of cells probed with the S1P3/Gαq/11 antibody pair are shown (A and B). Scale bar, 20 μm. Each bar graph shows quantification of at least 3 independent experiments as PLA signal (red dots) per cell (blue nuclei) relative to the control condition, with error bars indicating SE; n = 6 for C, n = 5 for A, B, E (Flow), and F (Flow), n = 3 for D, E (S1P), and F (S1P). *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: Human coronary artery endothelial cells (HCAECs) from male donors were obtained from either Cell Applications (San Diego, CA) or Lonza (Walkersville, MD) and maintained in complete endothelial growth medium (EGM-2; Lonza) supplemented with 10% heat-inactivated FBS and penicillin-streptomycin.

Techniques: Activation Assay, In Situ, Proximity Ligation Assay