Figure 1.

Role of individual nucleotides on cell migration. HCLEs grown to confluence on 8-well glass-bottom chambers were serum starved for 16 hours. Scratch wounds were made using a gel loader tip, and medium was immediately aspirated and replaced with medium alone (Neg. Cont) or medium containing one of the following ligands (100 μmol/L ADP, 100 μmol/L ATP, 100 μmol/L UDP, 100 μmol/L UTP, or 0.5 nmol/L EGF). Positive controls were cultures that were wounded and in which the wound media were retained for the duration of the time course [wound media (WM)]. Live cell migration was monitored using a laser-scanning confocal microscope equipped with an environmental chamber that maintained the cell environment at 37°C and 5% CO₂. Images were taken at each location every 20 minutes for 16 hours. Wound closure was determined using the region of interest function of the LSM software version 4.0, and percentage wound closure was calculated. Wound closure is indicated by the average closure at each time point ± SEM. Data are representative of three independent experiments.