Adenine nucleotide translocator 2 shRNA induced cytotoxic bystander effects on MDA-MB-231 cells. To evaluate bystander effects, MDA-MB-231 cells were transfected with adenine nucleotide translocator (ANT) 2 shRNA-1, shRNA-2 and shRNA-3. After 48 hours of incubation, nontransfected cells were cultured with the media obtained from ANT2 shRNA-1, shRNA-2 or shRNA-3 transfected cells for the next 24 hours. These nontransfected cells were then stained with annexin V–fluorescence isothiocyanate (FITC) and propidium iodide (PI) for flow cytometric analysis. Data are representative of three independent experiments.