Figure 5.
Effects of hepcidin on ferroportin expression and subcellular localization in BMDMs. (A) BMDMs were untreated (-) or treated (+) with human hepcidin (25 amino acids [aas]) for 3 hours. (B) Macrophages were untreated or treated for 6 hours with Fe-NTA in the presence or in the absence of human hepcidin. Postnuclear cell extracts (10 μg/lane) were then analyzed by Western blot using antiferroportin (Fpn) or anti–β-actin (loading control) antibodies. The position and size in kilodaltons of molecular mass markers are indicated on the right. (C) Immunofluorescence staining of ferroportin in control cells and in cells pretreated with Fe-NTA (16 hours, 100 μM) in the absence or presence of human hepcidin for the indicated period of time.