FIG. 10.

ROS are important mediators of H-1PV cytotoxicity in HeLa cells. (A) Twenty-four hours after infection with H-1PV (MOI, 1 PFU/cell), HeLa cells were loaded with DCFH-DA (10 μM, 1 h) and analyzed by FACS for ROS production. ROS accumulation was neutralized by the addition of the ROS scavenger GSH (5 mM). (B) Twenty-four and 48 h after infection, cell lysates were prepared and analyzed by Western blotting with the indicated antibodies. (C) Cell cycle distribution (top) and percentage of sub-G₁ apoptotic cell population (bottom) of mock-treated versus H-1PV-infected cells grown in the presence or absence of GSH (5 or 10 mM). A minimum of 20,000 cells were acquired and analyzed by FACS as described in Materials and Methods. Bars represent the means with relative standard deviations from three independent experiments, each performed in triplicate.