Jc 1 staining dye assay kit

ROS were determined by the Reactive Oxygen Species Assay Kit (Solarbio, China). The Δψm was determined using a JC-1 Staining Dye Assay Kit (Solarbio, China) according to the manufacturer’s instructions. Briefly, the cells were observed by fluorescence microscopy and analyzed by flow cytometry. The experimental details are provided in the Supplemental Methods.

The changes in mitochondrial membrane potential (Δψm) were observed with JC-1 staining dye assay kit (Beijing Solarbio Science & Technology Co., Ltd., Beijing, China), according to the method described by early literature [86 (link)]. JC-1 dye is a lipophilic cationic fluorescent dye with dual (red and green) emission wavelengths, and the attenuation of its red/green fluorescence intensity ratio is commonly used to represent an increase in mitochondrial membrane depolarization. The cells were cultured on the confocal dish with different treatments. Then, the old medium was replaced with fresh medium containing JC-1. Cells continued to incubate at 37 °C for 25 min and were then washed twice with ice-cold PBS. Cell morphology and staining were observed and photographed by an inverted fluorescence microscope (ZEISS Axio Observer A1, Oberkochen, German) and the intensities of fluorescence were analyzed by the ImageJ software (version No. 1.53e).