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. 1999 Mar 1;189(5):831–841. doi: 10.1084/jem.189.5.831

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Colocalization of Nramp2 and transferrin in early endosomes was determined by double immunofluorescence and confocal microscopy in RAW macrophages transfected with a c-myc–tagged Nramp2 (A–C) and in untransfected control cells (D–F). Cells were cultured in the presence of FITC–conjugated transferrin before fixation and immunostaining with the primary anti–c-myc tag antibody (9E10) and a secondary rhodamine-conjugated, anti–mouse antibody. The slides were then examined by confocal microscopy, and the FITC (green; B and E) and rhodamine (red; A and D) images were overlaid to identify colocalization (C and F). The image in C shows colocalization of Nramp2 and FITC–transferrin in several of the cells in the field.