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. Author manuscript; available in PMC: 2009 Jan 1.
Published in final edited form as: Blood Cells Mol Dis. 2007 Oct 1;40(1):132–138. doi: 10.1016/j.bcmd.2007.07.009

Figure 2. Pulse-chase study of hepcidin processing in Hep-G2 cells treated with and without furin proproteinase inhibitor.

Figure 2

Cells were labeled with 35S-met-cys for 1 hr then subjected to cold chase for the times in minutes indicated above the lane. Lane EQ was labeled for 3 hours without cold chase prior to processing to incorporate radioactivity into all peptide forms. Cell lysates (top panel) or the corresponding culture media (lower panel) were immunoprecipitated with either anti-pro (propiece) or anti-mature (hepcidin) antibody and analyzed on SDS-tricine PAGE. The autoradiograms of the gels are shown with the molecular weight standards indicated on the right side of the figure. Three forms of hepcidin are seen as marked by arrows. A cleaved form of the propiece is indicated by the small arrow (p) and the asterisk (panels B and D). In the right panel, cells were treated with furin inhibitor (decanoyl-RVKR-CMK) during the amino acid depletion step (1 hr) and during the radiolabeling procedure.