ROS generation and DNA damage accumulation in carnosol-treated MDA-MB-231 cells.

<p>(<b>A</b>) Carnosol induces ROS generation in MDA-MB-231 cells. Cells were treated with indicated concentrations of carnosol for 24 h and ROS production was analyzed, as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0109630#s2" target="_blank">Material and Methods</a>, using DCFDA fluorescence stain. (<b>B</b>) time-course measurement of ROS generation in carnosol-treated MDA-MB-231 cells. Cells were treated with 100 µM carnosol and ROS generation was examined at different time-point (1, 3 and 6 h). (<b>C–D</b>) Concentration and time-dependent accumulation of γH2AX, marker of DNA damage, in carnosol-treated cells. DNA damage was evaluated by measuring, by Western blotting, the level γH2AX accumulation using anti-phospho-H2AX (ser 139) antibody.