Fig. 1.

Overexpression of mZip14 increases iron and zinc accumulation by HEK 293H cells. (A) HEK 293H cells were transfected with either empty vector (pCMV-Sport6) or the same plasmid containing mZip14 and grown for 48 h. Western blots of total cell lysates (15 μg of protein per lane) were prepared by using SDS/PAGE and immunoblotting with anti-mZip14 antibody. (B) Time course of iron accumulation measured with ⁵⁹Fe in medium containing 2 μM ferric citrate (pH 7.4). (C) Time course of zinc accumulation measured with ⁶⁵Zn in medium containing 2 μM ZnCl₂ (pH 7.4), transfected with mZip14 (●) or vector alone (○). Nonspecific ⁵⁹Fe and ⁶⁵Zn binding to the cell surface was estimated by measuring uptake at 4°C. (D) Effect of BPS at 2.5- and 20-fold molar excess on mZip14-dependent iron accumulation after 60 min. Values are mean ± SD (n = 3; ∗, P < 0.05 compared with control) and are representative of multiple experiments.