Effect of FFA treatment on iron metabolism in HepG2 hepatocytes.

HepG2 hepatocytes were incubated with PA (0.66 mM) + OA (1 mM) or with FAC (150 μM) alone or in combination with FFA for 24 hours. A) Cells after Red Oil O staining. Intensity of red color is proportional to lipid accumulation. B) Absorbance of Red Oil O (540 nm). C) Intracellular iron concentration was measured by atomic absorption spectrometry. D) Ferritin H protein levels were evaluated by Western Blotting (lower part). Densitometric analysis of ferritin H protein levels is shown in the upper part of the figure; β-actin is shown as the loading control. E) TfR-1 mRNA levels evaluated by qRT-PCR. F) TfR-1 protein levels were evaluated by Western Blotting (lower part). Densitometric analysis of TfR-1 protein levels is shown in the upper part of the figure; β-actin is shown as the loading control. Results are mean values of three independent experiments, each experimental condition was evaluated in triplicate. Values are expressed as means±SD. AU, arbitrary units. *p<0.05 vs. controls.