Mitochondrial Complex I Inhibitors and Forced Oxidative Phosphorylation Synergize in Inducing Cancer Cell Death
Figure 3
FSK treatment enhances the viability loss induced by rotenone alone in MDA-MB-231 cells. MDA-MB-231 cells were cultured in 1 mM glucose and treated with 3 nM rotenone, 10 M FSK, or both molecules at 48 hours of culture. Cells were pretreated for 1 hour with FSK and then rotenone was also added for 4 hours, as shown in Figure 2(c). After treatment different parameters were investigated in untreated and treated cells. Viable cell count was performed using Trypan Blue (a). Percentage of cell death was evaluated after staining with PI and Annexin V-FITC (AnnV). Cells positive for one or both molecules were considered dead cells (b). After treatment cells were plated in normal growth medium for clonogenic assay and after ≥12 days colonies were stained and counted as reported in the histogram (c). (d) Intracellular ATP levels and (e) mitochondrial potential were also measured. All data represent the average of at least three independent experiments (±s.d.); , , (Student’s -test). (f) Basal OCR of MDA-MB-231 cells, grown 1 mM glucose and treated or not with FSK for 2–4 hours, was determined by Seahorse XF24 analyzer. Analysis was performed in three independent experiments (total number of samples ≥ 15) and values are indicated as the average ± s.e.m.; (Student’s -test).
