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Figure 2. Se uptake as a function of time and Zn2+ concentration in oocytes and mouse cell cultures; localization of ZIP8 to plasma membraneA. Se content following addition of Zn2+ (500 μM), or Zn2+ + HCO3− (3.5 mM) in selenite-treated Xenopus laevis oocytes expressing mouse ZIP8 cDNA (closed bar) versus control oocytes carrying vector only (open bar). Without added HCO3−, note that a basal level of HCO3− exists in the ND96 medium due to pCO2 in solution, as can be calculated by the Henderson-Hasselbach equation [30]. B. Se uptake kinetics in stably-transfected high-ZIP8-expressing ZIP8-MEFs vs control low-ZIP8-expressing LUC-MEFs during 20-min exposure to 50 μM HSeO3− + 100 μM Zn2+. C. Intracellular Se content in ZIP8-MEFs as a function of HSeO3− concentration; cells were treated with Zn2+ (100 μM) plus 5 to 100 μM HSeO3−. D. Intracellular Se content in ZIP8-MEFs (treated with 50 μM HSeO3− for 20 min) as a function of Zn2+ concentration. E. Intracellular Se content as a function of HSeO3− concentration (20-min treatment) in wild-type (WT) vs Slc39a8(neo/neo) knockdown mouse primary embryonic fibroblast (MEF) cultures; Zn2+ (100 μM) and HCO3− (25 mM) were constant. F. confocal microscopy of co-localization of ZIP8 (green) with the membrane marker Na+/K+-ATPase subunit (red) in ZIP8-MEF versus LUC-MEF cultures. DAPI, blue stain for DNA, i.e. nucleus. For all mouse culture experiments, HCO3− was present in culture medium at 25 mM. Addition of the HCO3− concentrations used in these experiments in either frog oocytes or mouse cultures did not significantly alter pH (7.5) of the medium. Brackets denote S.D. *P <0.05, **P <0.01. Image published in: McDermott JR et al. (2016) Copyright: © 2016 McDermott et al. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View

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