Fig. 2

Levels of NO and ROS induced by HZ in BMDM. BMDM were primed with IFN-γ for 2 h, and then treated with medium or different concentrations of HZ (12.5, 25, 50, 100 µM). Levels of nitrite (vertical axis) released into the supernatants after 24 h were measured. ^*P < 0.01, ^{*** #}P < 0.0001 vs control medium. Data displayed as averages ± SD of three independent experiments (a). BMDM were primed or not for 2 h. Cells were then stimulated with HZ (25 or 100 µM), MDP (2 µg/ml), or tert-butyl hydroperoxide (t-BHP) (10 µM). After each time point (0.5, 24 or 48 h) the supernatants were discarded and the cells were treated with 2′,7′-dichlorofluorescein diacetate (DCFDA) (15 µM) for 30 min at 37 °C. The cellular oxidative stress induced in macrophages was measured using the Synergy4 (BioTek) fluorescent microreader and the fluorescent units (FU, vertical axis) were recorded and correlated to the levels of ROS produced. ^#P < 0.0001 vs control medium. Data represent averages ± SD of three independent experiments (b)