Raw red channel MitoSOX fluorescence signal was plotted over time for each of the five groups using a selected ROI (see Materials & Methods). For consistency and reproducibility, six fields of view (FOVs) were imaged for cells in each treatment group, as well as the positive control and control groups. Figure 3a depicts raw fluorescence intensity for each group (n = 6), indicating the time points where MitoSOX Red indicator and drug treatments were added. After administering treatment, an increase in intensity was observed in all groups, implying a surge in the superoxide anion production rate. Additionally, as time increased, treatment groups showed a plateau or gradual increase in slope, validating the chosen duration of 8 hours.