Fig. 1
The presence of FPN in the SCV of Raw264.7 cells. a Raw264.7 cells pretreated with FeSO4 (0.1 mM) and/or hepcidin (1 μg/ml) for 3 h were infected with Salmonella (MOI, 100) and examined by confocal microscopy. Enlarged images of merge panels are shown in Supplementary Fig. 17. b The cells pretreated with PBS or hepcidin were infected with magnetized Salmonella (MOI, 100) for 2 h, and then SCV were isolated and Salmonella were analyzed by confocal microscopy. Staining for FPN is shown in green, and Salmonella in red. The isolated SCV were visualized by differential interphase contrast (DIC) imaging (scale bar 10 μm). Images are representatives of over 500 isolated SCVs per group from at least three independent experiments. c Raw264.7 cells pretreated with hepcidin were infected with magnetized Salmonella. FPN, Rab5, and annexin V in the SCV and the non-SCV fraction (supernatant) were examined by western blotting using specific antibodies. d Alternatively, Raw264.7 cells infected with Salmonella were treated with hepcidin for 2 h post infection and analyzed as in c
