Figure 3.

rabip4′ is associated with early endosomes. HeLa cells were transiently transfected with VSV-G-tagged rabip4′. To label the TfR pathway, cells were incubated for 30 min at 37°C with 25 μg/ml Alexa594-Tf (A-C). The cells were fixed and subjected to confocal immunofluorescence microscopy. rabip4′ was labeled with a mAb against the VSV-G tag and counterstained with Alexa488-labeled goat anti-mouse IgG (A and C) or Cy3-labeled goat anti-mouse IgG (D, F, G, and I). TGN46 detection was with a sheep antibody and Alexa488-goat anti-sheep (E and F), whereas LAMP-1 was detected with a rabbit antibody against its cytoplasmic tail and stained with Alexa488-goat anti-rabbit IgG (H and I). Merged images are shown in C, F, and I. Bar, 10 μm.