IS increases ROS production in LPS treated J774A.1 macrophages.

<p>ROS formation was evaluated by means of the probe 2′,7′ dichlorofluorescein-diacetate (H2DCF-DA) in J774A.1 cells. IS (500–250 µM) was added 1 h before and simultaneously to LPS (1 µg/ml) for 15 minutes. Panel A reports representative histograms for ROS detection. In panel B, ROS production was expressed as mean±s.e.m. of mean fluorescence intensity six independent experiments. In the same experimental conditions we also evaluated superoxide production by mitochondria adding MitoSOX Red. Panel C reports representative histograms for mitochondria superoxide detection. In panel D, mitochondrial superoxide production was expressed as mean±s.e.m. of mean fluorescence intensity of at least three independent experiments. Data were analyzed by analysis of variance test, and multiple comparison were made by Bonferroni's test. *** and ** denote P<0.001 and P<0.01 respectively versus LPS; °°°denotes P<0.001 versus control.