Ethidium bromide etbr

The mitochondrial function was examined using fluorescent probes. Cells were incubated with the probes for 30 min at 37 °C and then imaged using a BZ-X710 all-in-one fluorescence microscope (KEYENCE, Osaka, Japan). We used 10 μM MitoROS (mtROS) (10 μM, AAT Bioquest Inc., Sunnyvale, CA, USA) and 10 μM dihydrorhodamine 123 (DHR; Sigma-Aldrich, St. Louis, MO, USA) to assess oxidative stress, 100 nM MitoGreen (PromoCell GmbH, Heidelberg, Germany) to assess mitochondrial volume, 200 nM tetrathylrhodamine ethyl ester (TMRE; Sigma-Aldrich) to assess mitochondrial membrane potential, and 10 μg/mL ethidium bromide (EtBr; WAKO, for 10 min at room temperature) to assess mitochondrial DNA.

Ethyl methanesulfonate (EMS, CAS No. 62–50-0; Wako Pure Chemical Industries, Ltd., Tokyo, Japan) was used as a genotoxic positive control substance [14 ]. Physiological saline (PS) was used as a solvent (Otsuka Pharmaceutical Factory, Inc., Tokyo, Japan). Low melting agarose (LMA, NuSieve GTG) was purchased from Lonza (Basel, Switzerland), and standard melting agarose (SMA, Ultra Pure™ Agarose) was purchased from Thermo Fisher Scientific, Inc. (Massachusetts, USA). SYBR^Ⓡ gold was purchased from Thermo Fisher Scientific, Inc. (Massachusetts, USA). Ethidium bromide (EtBr) was purchased from Wako Pure Chemical Industries, Ltd. (Tokyo, Japan).