Fig. 4.
The effect of RelB on the induction of ferroptosis. (A, B) The levels of ROS in RelB-manipulated MCF7 cells were measured using a H2DCFDA probe with CDCFDA normalization (A). TAM-induced ROS in each cell line was plotted (B) (n = 4). (C) TAM-induced lipid peroxidation in RelB-manipulated cells was imaged by confocal microscopy with a BODIPY 581/591 probe. (D) After TAM-treatment, amounts of lipids and oxidative lipids in RelB-manipulated cells were measured by spectrophotometer. The ratio of oxidized/reduced BODIPY is indicated in a bar graph (n = 4). (E) TAM-induced lipid peroxidation in RelB-manipulated cells was confirmed by flow cytometry. (F) Furthermore, lipid peroxidation generated in RelB-manipulated cells was validated by MDA assay (n = 4). (G) The contents of iron ion in RelB-manipulated cells were measured using an iron colorimetric assay (n = 4). Data are shown as mean ± SD, * (p < 0.05) and **(p < 0.01); two-way ANOVA.