Fig. 2.

P2X7-induced ROS formation in MEL cells is impaired by NAC and DPI. H₂DCFDA-loaded MEL cells (left) or MEL cells (right) in NaCl medium were pre-incubated at 37 °C in the a, f absence (control) or presence of a 10 mM NAC for 30 min or f 1 mM l-NAME for 60 min or in the presence of b DMSO or 20 μM DPI for 30 min, c DMSO or 100 μM apocynin for 60 min, d DMSO or 5 μM rotenone for 60 min or e NaOH or 100 μM allopurinol for 30 min. a–f Cells were then incubated in the absence (basal) or presence of 1 mM ATP at 37 °C for 15 min; 25 μM ethidium⁺ was also present (right). Incubations were stopped by addition of MgCl₂ medium and centrifugation, and the mean fluorescence intensity (MFI) of DCF (ROS formation; left) or ethidium⁺ uptake (pore formation; right) analysed by flow cytometry. Results are mean ± SD (n = 3); *P < 0.05 compared with corresponding basal; **P < 0.01 compared with corresponding basal; ††P < 0.01 compared with ATP without NAC or antagonist; ‡‡P < 0.01 compared with basal control