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. Author manuscript; available in PMC: 2010 Mar 1.
Published in final edited form as: Cell Metab. 2009 Mar;9(3):217–227. doi: 10.1016/j.cmet.2009.01.010

Figure 6. The α3 and cytoplasmic domains of HFE mediate the Tf induction of hepcidin expression in HepG2 cells.

Figure 6

A. Schematic representation of the HFE chimeras including α1α2, α3tmcd, α3cd and cd constructs. HFE is shown in gray, HLA-B7 is shown in white. HFE chimeras were constructed by replacing the appropriate segments between the parent proteins. Individual recombinant proteins used throughout the study are referred to by the acronyms to left of the construct cartoon. B. Generation of HFE-HLA-B7 chimeras. HFE chimeras including stable cells lines HepG2/tTA α1α2, HepG2/tTA α3tmcd, HepG2/tTA α3cd and HepG2/tTA cd were generated as described in Experimental Procedures. Lysates (25 μg) of cells, uninduced (−dox) or induced (+dox) were used to screen the positive clones by immunoblot using anti-FLAG antibody. C. The α3 and cytoplasmic domain of HFE is required for HFE-mediated Tf induction of hepcidin expression in HepG2 cells. All HepG2/tTA control and HFE chimeras cells were cotransfected with pLuc-link-HAMP and pCMV-β-gal, treated, and analyzed as described in Experimental Procedures. Results are expressed as average ± S.D. of three independent experiments performed in triplicate. P-values < 0.001 are indicated by *** and < 0.01 by **.