2.5.3. Intracellular ROS generation after UV irradiation
The dichlorofluorescin diacetate (DCFH2-DA) assay was used to assess the extent of ROS generation following UVA or UVB exposure. Cells were seeded in 12-well plates and incubated with tested extracts for 24 h. Following UV exposure, cells were washed twice with PBS and incubated with various concentrations of the extracts for 2 h. Following incubation cells were washed with PBS and incubated with 10 μM dihydrodichlorofluorescein diacetate (DCFH-DA) for 30 min at 37 °C. Cells were then detached using accutase, washed and fluorescence was determined using a FACS Calibur flow cytometer (Becton Dickinson, USA).