Figure 3.

The basal ROS level in A375, H9C2 and NHDF. (A,B): A375, H9C2 and NHDF were treated with 0.1 µM H₂DCF-DA and the level of DCF as a measure of intracellular ROS was determined by flow cytometry using a CyFlow Cube 6. FlowJo software was used to create a histogram overlay (A) and to calculate the MFI (B). (A) Representative histograms for treatment with H₂DCF-DA (dark histograms) were depicted, and unstained cells served as the negative control (light histograms). Fluorescence intensity is plotted against the cell number. (B) The MFI of the cells treated with H₂DCF-DA is depicted. The MFI of unstained cells served as the negative control and the MFI of A375 cells was set to 100%. Data represent means ± SEM, n ≥ 3. A one-way ANOVA was used to determine statistical significance compared to the control. * p ≤ 0.05, ** p ≤ 0.01.