Transferrin–iron incorporation in HC macrophages transfected with HFE.

(A) HC (solid bars) and normal (empty bars) macrophages were incubated for 24 hours in the presence of ⁵⁵Fe-Tf and cell-associated radioactivity measured at the indicated time points. Data are mean ± SD from 6 separate experiments performed in triplicate.^*P < .001 versus HC macrophages. (B) HC macrophages transduced with the pCMV-HFE (solid bars) or the pCMV-βgal (empty bars) plasmids were incubated for 24 hours in the presence of ⁵⁵Fe-Tf. Cell-associated radioactivity was measured at the indicated incubation time points. Data are mean ± SD of experiments in macrophages from 3 subjects with HC (A, B, C) performed in triplicate, representative of 8 separate experiments in HC macrophages. ^*P < .001 versus HC macrophages transduced with control vector. (Inset) Time-course of intracellular⁵⁵Fe-ferritin accumulation in HC macrophages transduced with the pCMV-HFE or the pCMV-βgal plasmid as evaluated by nondenaturing PAGE of pooled cell lysates coupled with⁵⁵Fe-autoradiography. Ft, ferritin.