Fig. 5.

PAP7 and DMT1 interact in BBM vesicles (BBMV) from rat duodenum. As described in the materials and methods, BBMV were isolated from duodenal epithelium cells of heterozygous Belgrade rats (+/b). Detergent-solubilized BBMV were passed through the columns of anti-PAP7 IgG and control IgG cross-linked to agarose beads as described in materials and methods. After being extensively washed, proteins were eluted from the beads and the eluates and an aliquot of the BBMV were separated by SDS-PAGE followed by Western blot analysis with anti-DMT1 [iron-responsive element (IRE)]. Subsequently, the antibodies were removed from the same membrane as described in the materials and methods and Western blot analysis repeated using anti-PAP7 antiserum.