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Figure 8


Fig. 8. Altered processing of the juvenile hemochromatosis-linked RGMc G313V mutation. (A) Detection of HA-RGMc (WT) and HA-RGMc G313V in whole-cell protein extracts (WCE) and conditioned culture medium (CM) in transiently transfected in Cos-7 cells. Full-length and internally cleaved RGMc proteins are indicated by black or gray arrows, respectively. (B) Detection of RGMc WT and G313V proteins after labeling of cell membranes with the EZ-link reagent and streptavidin-agarose pull down. Immunostaining for tubulin and cadherin (lower panels) is included for loading and cell-surface specificity controls, respectively. (C) Immunostaining with anti-HA antibody of samples from B. In B and C, the full-length, internally cleaved and anomalously processed RGMc proteins are indicated by black, gray or open circles respectively.





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