Figure 4.

VE dietary supplementation reduced intracellular ROS levels in TGFα/c-myc hepatocytes. Hepatocytes were isolated by collagenase perfusion as described in Materials and Methods, incubated in the absence or presence of 5 μM of the oxidative-sensitive probe DCFH-DA for 30 min at 37°C and subjected to flow cytometry. The figure represents the overlay of three representative fluorescence-activated cell sorter analysis performed on TGFα/c-myc hepatocytes isolated from 10-wk-old mice fed with control (gray area) or VE-supplemented diet (black area). The white area shows the autofluorescence profile in unstained TGFα/c-myc hepatocytes. DCF fluorescence was measured as difference between DCF fluorescence and autofluorescence in unstained cell suspensions for each cell preparation analyzed. Representative result from three independent experiments is shown.