Subcellular localization of Nramp2/DMT1 in transfected RAW macrophages.

Immunofluorescence detection of a c-Myc–tagged Nramp2 protein expressed in RAW macrophages using a mouse monoclonal anti–c-Myc antibody (9E10) and a Cy3-labeled secondary antimouse antibody (red staining). The recycling endosome marker EEA1 was detected using a goat antiserum revealed with Alexa 488–coupled secondary antibody (green staining, left panel A). The late endosomal/lysosomal marker, Lamp1, was detected with a specific rat monoclonal antibody and revealed with FITC-conjugated antirat antibody (green staining, right panel B). Cells were examined under a × 63 objective by confocal microscopy. Green staining was overlaid with the red staining (yellow shows colocalization).