Author: Dong, Xiaoxv; Ni, Boran; Fu, Jing; Yin, Xingbin; You, Longtai; Leng, Xin; Liang, Xiao; Ni, Jian
Title: Emodin induces apoptosis in human hepatocellular carcinoma HepaRG cells via the mitochondrial caspase-dependent pathway Document date: 2018_8_2
ID: 02r8i5n1_8
Snippet: Determination of intracellular ROS. ROS generation was measured with an oxidation-sensitive probe DCFH-DA, which is cleaved by nonspecific esterases and yields highly fluorescent 2,7-dichlorofluorescein (DCF) upon oxidation by ROS. After treatment with emodin (20, 40 and 80 µM) or control medium treatment for 24 h, the cells were washed twice in PBS. Then, they were loaded with DCFH-DA (10 µM) and incubated for 30 min at 37˚C in the dark (34) .....
Document: Determination of intracellular ROS. ROS generation was measured with an oxidation-sensitive probe DCFH-DA, which is cleaved by nonspecific esterases and yields highly fluorescent 2,7-dichlorofluorescein (DCF) upon oxidation by ROS. After treatment with emodin (20, 40 and 80 µM) or control medium treatment for 24 h, the cells were washed twice in PBS. Then, they were loaded with DCFH-DA (10 µM) and incubated for 30 min at 37˚C in the dark (34) . Thereafter, the cells were harvested and washed twice with PBS, and the oxidation-induced increase in DCF fluorescence was subsequently measured by flow cytometry.
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