P2X7 Receptor Activation Induces Reactive Oxygen Species Formation and Cell Death in Murine EOC13 Microglia
Figure 3
EOC13 microglial cells express functional P2X7. (a and b) EOC13 cells in NaCl medium were incubated with 25 μM ethidium+ in the absence (basal) or presence of (a) 1 mM ATP, 0.1 mM BzATP, or (b) varying concentrations of ATP (as indicated) at 37°C for 5 min. (c and d) Cells in NaCl medium were preincubated in the absence (control) or presence of (c) 30 μM Brilliant Blue G (BBG), 100 μM A438079, 30 μM AZ11645373, or (c and d) 10 μM AZ10606120 at 37°C for 15 min. (c) Ethidium+ (25 μM) or (d) YO-PRO-12+ (1 μM) was then added, and (c and d) cells were incubated in the absence (basal) or presence of 1 mM ATP at 37°C for 5 min. (a–d) Incubations were stopped by the addition of MgCl2 medium and centrifugation. Mean fluorescence intensity (MFI) of fluorescent cation uptake (pore formation) was determined by flow cytometry. (a, c, and d) Results shown as means ± SD, ; *** compared to corresponding basal; ††† compared to corresponding ATP in the absence of antagonist. (b) Curve presented as a percentage of the maximal ATP-induced ethidium+ uptake and expressed as the mean ± SD, .
