Fig. 2. Stimulation of S. cerevisiae acetic-acid-induced apoptosis by expression of mammalian PKC-, - or - but not PKC- isoform is associated with significant ROS production. ROS production by mitochondria was detected by flow cytometry, using MitoTracker Red CM-H₂XRos. Overlays of red fluorescence histograms were obtained with yeast treated with 0 and 300 mM acetic acid: black fill, yeast transformed with the empty vector (YEplac181 or YEp51); light-grey line, yeast expressing PKC-, -, - or -. Cells treated in the absence of acetic acid (0 mM) were used as a negative control. Data represent one of two independent experiments.