Figure 3

Localisation of hepcidin protein in rat brain endothelial cells. Double immunofluorescence (IFC) staining was performed with polyclonal rabbit anti-hepcidin and other monoclonal antibodies with counterstaining for nuclei with DAPI (Blue). Hepcidin protein was expressed in epithelial cells of the choroid plexus where minimal co-localisation with the pericyte marker PDGFβR1 was observed (arrow in panel A-C). At higher magnification hepcidin was visible in the endothelial lining of blood vessels where it co-localised with the endothelial marker CD31 (D-F), but not with the glial marker chondroitin sulphate proteoglycan neuron ⁄ glia antigen 2 (NG2) (G-I). At higher magnification hepcidin protein was visible in blood vessel walls (H-I). Hepcidin protein was seen in the corpus callosum with some co-localisation with ferroportin noted (J-L), and a similar finding was seen in a section of third ventricle (M). Hepcidin was present in the vascular endothelium of the circumventricular organs where co-localisation with VEGF was seen (N). In the sub-ventricular zone hepcidin was co-localised with S100β (O). Protein expression and co-localisation was indicated with arrow in relevant panels. The scale bar in O represents 70 μm in A and B and M; 20 μm in D-F, H and I; 50 μm in G; 30 μm in C, J-N and O.