Figure 5
A. Iron loading increases expression of RGMc on the cell membrane and diminishes accumulation in extra-cellular fluid. A. Hep3B cells infected with Ad-HA-RGMc were incubated for 24 h with the concentrations of ferric ammonium citrate (FAC) indicated, followed by cell-surface biotin labeling, and detection of RGMc by immunoblotting in cell lysates (left panel), on the membrane after surface biotin labeling and streptavidin pull-down (middle panel), and in the medium (right panel). Similar results were observed with Cos-7 cells. B. Detection of RGMc by immunoblotting of conditioned medium from transiently transfected Cos-7 cells following incubation for 24 h with the concentrations of FAC or deferoxamine (DFO) indicated. Identical results were seen with Hep3B cells. For A – B, arrows are as in legend to Fig. 2.