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. 2000 Feb;11(2):747–763. doi: 10.1091/mbc.11.2.747

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Copyright © 2000, The American Society for Cell Biology

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Immunoelectron microscopy for GFP-SKD1^E235Q, the endogenous SKD1, TfR, and EEA1. HEK293 cells transfected with GFP-SKD1^E235Q were fixed, and the localization of GFP-SKD1^E235Q (a), TfR (c), and EEA1 (e–h) was examined by silver-enhanced immunogold electron microscopy using antibodies against GFP, TfR, and EEA1, respectively. Distribution of the endogenous SKD1 in H-4-II-E cells was also examined using antibodies against SKD1 (b). (d) Double stain of GFP-SKD1^E235Q (silver-enhanced gold particles) and TfR (HRP-DAB staining). V, exaggerated aberrant multivesicular vacuole; T, tubulo-vesicular structures. Arrows in b, endogenous SKD1 associated with membrane compartments/vesicles; arrowheads in f–h, EEA1 associated with protrusions of the vacuoles. Bar, 1 μm..

Immunoelectron microscopy for GFP-SKD1^E235Q, the endogenous SKD1, TfR, and EEA1. HEK293 cells transfected with GFP-SKD1^E235Q were fixed, and the localization of GFP-SKD1^E235Q (a), TfR (c), and EEA1 (e–h) was examined by silver-enhanced immunogold electron microscopy using antibodies against GFP, TfR, and EEA1, respectively. Distribution of the endogenous SKD1 in H-4-II-E cells was also examined using antibodies against SKD1 (b). (d) Double stain of GFP-SKD1^E235Q (silver-enhanced gold particles) and TfR (HRP-DAB staining). V, exaggerated aberrant multivesicular vacuole; T, tubulo-vesicular structures. Arrows in b, endogenous SKD1 associated with membrane compartments/vesicles; arrowheads in f–h, EEA1 associated with protrusions of the vacuoles. Bar, 1 μm..