Intracellular reactive oxygen species (ROS) production was measured by epifluorescence using the described protocol [26 (link)]. Briefly, the Huh7 cells were seeded into 48-well plates, transfected with plasmids encoding HCV core variants. Cell culture medium was removed 30 h post transfection, and cells were incubated in the medium containing 25 µM 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA) for 30 min. The treated cells were washed with PBS, resuspended in PBS, and the fluorescence intensities were measured using Plate CHAMELEON V reader (Hidex Oy, Åbo, Finland) with excitation at 485 and emission at 535 nm.
Jansons J., Sominskaya I., Petrakova N., Starodubova E.S., Smirnova O.A., Alekseeva E., Bruvere R., Eliseeva O., Skrastina D., Kashuba E., Mihailova M., Kochetkov S.N., Ivanov A.V, & Isaguliants M.G. (2019). The Immunogenicity in Mice of HCV Core Delivered as DNA Is Modulated by Its Capacity to Induce Oxidative Stress and Oxidative Stress Response. Cells, 8(3), 208.
Corresponding Organization : Chumakov Institute of Poliomyelitis and Viral Encephalitides
Other organizations :
Latvian Biomedical Research and Study Centre, Engelhardt Institute of Molecular Biology, R.E. Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology, National Academy of Sciences of Ukraine
Fluorescence measurement (excitation at 485 nm, emission at 535 nm)
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