FIGURE 3

FIGURE 3

Iron supplementation can increase ROS level and reduce mitochondrial damage in cells under hypoxia stress. (A,B) Western blot analysis and quantification of Ferritin expression in ZFL cells cultured under normoxia and hypoxia treated with FAC, DFO and Fer-1 for 3 days. Actin as a loading control. (C–E) Analysis of changes in total ROS (C), lipid peroxidation level (D) and mitochondrial-derived ROS (E) levels in cells with H2DCFDA, C11-BODIPY and MitoSOX probe under normoxia and hypoxia for 3 days. Cells under hypoxic stress were rescued with FAC (2.5 mM), DFO (10 µM) and Fer-1 (2.5 µM). (F,G) Quantitative results and representative images of cellular JC-1 fluorescence in normoxic cell and hypoxic cells treated with FAC (2.5 mM), DFO (10 µM) and Fer-1 (2.5 µM) under hypoxia stress. Normoxia was used as a control group for significance analysis. Error bars, mean ± s.d., n = 3 (biological replicates).