Cellular Oxidative Stress Assessment

For cellular ROS detection, cells were incubated with 10 μM of 2,7-Dichlorodihydrofluorescein diacetate (DCFH-DA) (MedChemExpress, NJ, USA) working solution at 37°C for 20 min in darkness. The fluorescence light intensity was measured by using a multifunction microplate reader (Infinite 200 Pro, Tecan, Australia). Cellular glutathione (GSH), oxidative glutathione (GSSG), superoxide dismutase (SOD) activity, and malonaldehyde (MDA) levels were measured by using the corresponding commercial assay kits (Boxbio, Beijing, China), according to the manufacturer’s instructions.

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Zhou H., Wan S., Luo Y., Liu H., Jiang J., Guo Y., Xiao J, & Wu B. (2023). Hepatitis B virus X protein induces ALDH2 ubiquitin-dependent degradation to enhance alcoholic steatohepatitis. Gastroenterology Report, 11, goad006.

Publication 2023

DCFH-DA Infinite 200 Pro

Assay Cellular Darkness Fluorescence Glutathione Gssg Light Malonaldehyde Superoxide dismutase

Corresponding Organization : First Affiliated Hospital of Jinan University

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Variable analysis

independent variables

Incubation of cells with 10 μM of 2,7-Dichlorodihydrofluorescein diacetate (DCFH-DA) working solution

dependent variables

Fluorescence light intensity
Cellular glutathione (GSH)
Oxidative glutathione (GSSG)
Superoxide dismutase (SOD) activity
Malonaldehyde (MDA) levels

control variables

Incubation time (20 min)
Incubation temperature (37°C)
Incubation conditions (in darkness)

controls

No positive or negative controls were explicitly mentioned in the provided information.

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