FIGURE 1

FIGURE 1

Hypoxia stress affects cell growth, ROS production and iron metabolism in cytoplasma and mitochondria. (A) Microscopic analysis of morphological changes of ZFL cells under normoxia or hypoxia for 1, 2, 3 and 4 d. (B) Cell Viability analyzed with PrestoBlue™ HS Cell Viability Regent under normoxia or hypoxia for 1, 2, 3 and 4 d. (C–E) Analysis of changes in total ROS (C), mitochondrial-derived ROS (D) and lipid peroxidation (E) levels in cells with CM-H2DCFDA, MitoSOX and C11-BODIPY probe under normoxia and hypoxia for 3 days. (F) Western blot analysis of Ferritin expression in ZFL cells cultured under normoxia and hypoxia for 3 days. (G) Phen Green™ SK (PGSK) probe analysis and quantification of cytoplasmic free iron content in ZFL cells after treated under normoxia and hypoxia for 3 days. (H) The mRNA expression of mitochondrial iron storage gene fth31 in ZFL cells quantified by real-time RT–PCR under normoxia and hypoxia for 3 days. (I) Fluorescence microscope with RPA red indicator analysis and quantification of mitochondrial iron content in ZFL cells treated under normoxia and hypoxia for 3 days. Normoxia was used as a control group for significance analysis. Error bars, mean ± s.d., n = 3 (biological replicates).