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Figure 6


Fig. 6. Localization of Pmel17 in the plasma membrane. (A,B) Plasma membrane (PM) proteins recovered by crosslinking and SIG purification were analyzed by immunoblotting. All samples were processed in parallel with negative controls. Arrowheads indicate the fully glycosylated ~100 kDa band (white), the partially glycosylated 85 kDa band (black), and the cleaved 26 kDa band of Pmel17 (gray). The black arrow identifies a ~70 kDa band. (A) Pmel17 was recovered using a biotinylated reagent from the plasma membrane in M14 cells and in MNT1 cells. Recovery of the resident plasma membrane protein, I5{alpha}, was used as control for the extraction method (see bottom panel). (B) Plasma membrane fraction after SIG purification (left). Using immunoblot analysis, Pmel17 was detected with the {alpha}PEP13h antibody both in total extracts (TE) and in the SIG fraction. (C) Plasma membrane fractions were analyzed for the presence of markers as noted. Images are representative of two independent experiments.





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