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Detection of reactive oxygen species (ROS)
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ROS production was evaluated using 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA, Kodak)18. Fresh stock solution of 2 mM (w/v) DCFH-DA in ethanol was prepared and kept at − 20 °C in the dark until the use. Briefly, cells were incubated with DCFH-DA solution (10 µM) at 37 °C for 1 h in a humidified atmosphere with 5% CO2. After washing, the fluorescence of cells was acquired by FACS (i.e. λex 488 nm, λem 530 nm).
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