Fig. 7

Trichomonas vaginalis ESP induced apoptosis and ER stress response and generated mitochondrial ROS production in SiHa cells. SiHa cells were treated with 100 μg/ml T. vaginalis ESP for 0, 2 or 6 h. a, b Cellular ROS production was measured by confocal microscopy (a) and flow cytometry (b) with the CellROX oxidative stress reagent, a fluorogenic probe for measuring oxidative stress in live cells. Plots depict the CellROX-positive cells as determined by fluorescence analysis of flow cytometry. Asterisks indicate significant difference (***P < 0.001) in mean fluorescence compared with the untreated control cells under the same conditions. c, d Mitochondrial ROS production was determined by confocal microscopy (c) and flow cytometry (d) with MitoSOX, a mitochondrial ROS dye. Plots depict the MitoSOX-positive cells as determined by fluorescence analysis of flow cytometry. Data shown are representative of three independent experiments with similar results. Asterisks indicate significant difference (***P < 0.001) in mean fluorescence compared with the untreated control cells under the same conditions. e Apoptosis- and ER stress-related protein expression levels were evaluated by western blot. The data shown are representative of three independent experiments with similar results. Scale bars: 10 µm. Abbreviations: CTL, Untreated control SiHa cells; ESP, SiHa cells treated with T. vaginalis ESP