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. 2012 Aug;136(4):448–458. doi: 10.1111/j.1365-2567.2012.03601.x

Figure 4.

Figure 4

ATP induces reactive oxygen species (ROS) production in macrophages in vitro. (a) Peritoneal exudate macrophages (PEMs) were stimulated with 300 μm ATP for 15 min at 37° and then treated with 5 μm 5-(and-6)-chloromethyl-2′,7′-dichlorodihydrofluorescein diacetate, acetyl ester (CM-H2DCFDA) for 10 min at 37°. The PEMs were visualized immediately with a fluorescence microscope (objective lens × 20). (b) PEMs were stimulated with several concentrations of ATP for 5, 15 and 30 min and ROS production was measured using the fluorescent probe, CM-H2DCFDA. Data represent the mean ± SEM of > 400 cells (or for 3 mm ATP-stimulated PEMs: > 200 cells). One result from two representative experiments is shown. (c) PEMs were pre-incubated for 60 min with N-acetyl-l-cysteine (NAC) before the addition of 300 μm ATP. Stimulation in the presence of NAC continued for 24 hr. Data represent the mean ± SEM of triplicate samples. The experiments were repeated three times.