(A) Visualization of HT-1080 cell viability over time +/− erastin (Era, 10 µM) and deferoxamine (DFO, 100 µM). (B,C) Cytosolic and lipid ROS production assessed over time (2,4 and 6 hrs) by flow cytometry using H2DCFDA and C11-BODIPY. (D) Mitochondrial ROS assessed in HT-1080 cells treated for 6 hrs with erastin+/−DFO, as above, or with rotenone (250 nM)+/−DFO. In (A–D) representative data from one of four experiments is shown. (E) Erastin-induced death in 143B ρ0 and ρ+ cells. (F) mtDNA-encoded transcript levels in ρ0 and ρ+ cells. Results in (E) and (F) are mean+/−SD from one of three representative experiments.