H2O2 caused depolarization of mitochondrial membrane potential. Cells were treated with various concentrations of H2O2 for 0.5 hour (A), 1 hour (B), 2 hours (C) or 4 hours (D), then processed for the JC-1 assay as described in the Materials and Methods. Emission readings at 545 nm and 595 nm were plotted as a function of H2O2 concentration. Results from (A-D) were re-plotted and presented as (E) to illustrate a dose- and time-dependent increase of 545/595 emission intensity ratio. (F): Cells were treated with various concentrations of FCCP for 4 hours, then the 545/595 emission intensity ratio of each treatment was determined. Results indicated that FCCP caused a dose-dependent increase of the ratio, which reached a maximum at ~20 μM. Note the degree of mitochondrial depolarization caused by 2 mM H2O2 was similar to that caused by 10 μM FCCP. Results of H2O2 treatment were derived from 4 experiments (total 12 replicates), and those of FCCP treatment were from 3 experiments (total 9 replicates).
Garg and Chang BMC Pharmacology 2004 4:6 doi:10.1186/1471-2210-4-6