Figure 4
PSY induces mitochondrial ROS production. (a) Representative dot-plots of MO3.13 cells stained with Mitotracker Red CM-H2XRos and analyzed by flow cytometry. The side-scattered light signal (SSC) is also reported. The R6 region was set by the dot-plot obtained for the control population in (no PSY, 10% FBS). (b) Mitochondrial ROS production in case of cell starvation (0% FBS) and culture maintenance condition (10% FBS). (n=6, t-test). (c) PSY dose–response quantification of mitochondrial ROS production (n=6, one-way ANOVA (Dunnett’s post-hoc test ) PSY 3–10 μM versus Untreated). (d) Representative traces of mitochondrial ROS (orange line) and cytoplasmic Ca2+ (green line) kinetics upon PSY 10 μM administration (t=0; black arrow) during apoptotic cell death (n=3, >15 analyzed cells). PSY induced a first, rapid ROS production peak correlated with the first Ca2+ peak; a second, more protracted, mitochondrial ROS elevation was recorded before cell lysis (blue arrow). (e) Time-lapse images of the Mitotracker Red CM-H2XRos fluorescence of a representative cell is reported below the traces
