| Method |
Antibodies and Recombinant Proteins, Cell Culture of HeLa cells, Immunoblot Analysis, Immunofluorescence Microscopy, Antibody Internalization Assay, Whole-Mount Electron Microscopy, Immunoelectron Microscopy, Metabolic Labeling and Immunoprecipitation, Epidermal Growth Factor Receptor (EGFR) Down-Regulation, TIP47/Rab9 Binding Studies, Crude Membrane Fractionation. To explore the importance of Rab9 for microdomain establishment, researchers depleted the protein from cultured cells. Rab9 depletion decreased late endosome size and reduced the numbers of multilamellar and dense-tubule–containing late endosomes/lysosomes, but not multivesicular endosomes. |