Mitocapture mitochondrial apoptosis detection fluorometric kit

The mitochondrial membrane potential (MMP) was analyzed using a MitoCapture Mitochondrial Apoptosis Detection Fluorometric kit (BioVision, Milpitas, CA, USA). In brief, cells were collected from each treatment and centrifuged at 3000 rpm for 5 min, and then, the supernatant was discarded. Pelletized cells were gently suspended by adding 1 mL of MitoCapture solution and incubated at 28 °C for 20 min in the dark. After another centrifugation at 3000 rpm for 5 min, pelleted cells were re-suspended with 1 mL of incubation buffer and analyzed with a FACScan flow cytometer (BD Biosciences, San Jose, CA, USA).

A FITC Annexin V/PI apoptosis detection kit was purchased from BD Biosciences (NY, USA). Hoechst 33258 and PI dye detection kits were provided by Solarbio Life Science (Beijing, China). MitoTracker® probes and a Pierce^™ BCA protein assay kit were provided by Thermo Fisher Scientific (MA, USA). FITC-klawas provided by GL Biochem Ltd. (Shanghai, China). MitoCapture Mitochondrial Apoptosis Detection Fluorometric Kit was provided by BioVision(SF,USA). The human breast cancer cell line MCF-7, lung cancer cell line A549, and mouse embryonic fibroblast cell line NIH3T3 were obtained from the Institute of Biochemistry and Cell Biology, Shanghai Institute for Biological Sciences, Chinese Academy of Sciences (Shanghai, China).