Mitochondrial membrane potential assay kit with jc 1 c2006

Standard chemicals such as ZEN were purchased from Pribolab (Qingdao, China), of which the purity grade is more than 99%. TM4 cell line and HepG2 cell line were purchased from Pricella (Wuhan, China). Luria–Bertani agar (LB) plates were used to screen microbial cultures for ZEN biotransformation capability, isolate single colonies, and construct a culturable microbiome of rhizosphere soil of Pseudostellaria heterophylla. CM-0456 medium with FBS, penicillin, and streptomycin were used to culture the TM4 and HepG2 cell. An annexin V-FITC/PI double-staining cell-apoptosis detection kit (KGA108) was purchased from KeyGEN BioTECH (Nanjing, China). A mitochondrial-membrane-potential assay kit with JC-1 (C2006) and a reactive oxygen species assay kit (S0033) was purchased from Beyotime Biotechnology (Shanghai, China).

The chrysanthemum (chuju) extract was obtained by aqueous extraction. In brief, after drying, chrysanthemum was extracted at 80 °C for 5 min by adding deionized water at 1:20 (w/w). Extractions were performed three times in total. The raw extract was pooled and filtered through a 0.22 μm filter. The filtrate was then freeze-dried. L-O2 human hepatocyte was commercially acquired from Zhong Qiao Xin Zhou Biotechnology Co., Ltd. (Shanghai, China). Hydrogen peroxide (H₂O₂) was purchased from Hushi (Shanghai, China). Both the 3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide (MTT) cell proliferation and cytotoxicity assay kit (M1020) and 2′,7′-dichlorofluorescin diacetate (4091-99-0) were purchased from Solarbio Science & Technology Co., Ltd. (Beijing, China). The malondialdehyde (MDA) assay kit (TBA method, A003-1-2), superoxide dismutase (SOD) assay kit (WST-1 method, A001-3-2), glutathione peroxidase (GSH-Px) assay kit (A005-1-2), catalase (CAT) assay kit (A007-1-1) and annexin V cell apoptosis assay kit (G003-1-2) were purchased from JianCheng Bioengineering Institute (Nanjing, China). The cell cycle assay kit (propidium iodide staining method) was purchased from Zhi En biology (Hefei, China). The mitochondrial membrane potential assay kit with JC-1 (C2006) was purchased from Beyotime (Shanghai, China).