Nucleotides Regulate Secretion of the Inflammatory Chemokine CCL2 from Human Macrophages and Monocytes
Figure 1
Nucleotides induce intracellular calcium responses in THP-1 cells. (a) Representative plot showing P2Y2 expression on THP-1 monocytes using a specific anti-P2Y2 antibody and a BD FACSCalibur flow cytometer. (b) THP-1 cells were loaded with Fluo-4 using a NW kit and plated at 2 × 105 cells/well in poly-D-lysine coated 96-well plates. Intracellular calcium responses were measured using a BMG Labtech Fluostar Optima plate reader for over 60 seconds at 37°C. Responses to 100 nM ATP, 50 nM UTP, 50 nM 2-thio-UTP, and 250 nM UTPγS are shown in black. Paired responses of cells pretreated with the P2 receptor antagonist suramin (30 μM; 30 minutes) are shown in red. (c) Mean calcium response data from several experiments. (d) A concentration-response curve was generated for UTP-induced calcium responses on THP-1 cells. Mean responses ± SEM are plotted from 4 separate experiments.
