Deficiency of mitochondrial energy metabolism accompanies cell death in cells treated with MIPP. For all graphs the solid bars are values for cells treated with 10 μM MIPP and the open bars are controls treated with an equivalent volume of DMSO. A) ATP levels were measured as described in the Methods. The decrease in ATP in MIPP-treated cells on day-2 (*) was significant at p < 0.02 compared to the cells treated with DMSO. B) Mitochondrial membrane potential was evaluated using the JC-1 assay. The significant decline (p < 0.0001) in the ratio of fluorescence emission at 600 nm vs. 535 nm on day-2 signifies a disruption of mitochondrial membrane potential. C) There is no increase in intracellular ROS associated with MIPP-induced cell death, based on measuring the fluorescence of the ROS indicator, H2DCFDA. The decline in ROS in the MIPP-treated cells on day-2 was significant at p < 0.005.