Figure 7.

The FHF complex promotes timely trafficking of EGF through the endosomal pathway. (A) Fluorescently labeled EGF (EGF-TR, Texas Red) was used in endosomal trafficking assays as described in Materials and Methods. At either 5 min or 2 h after initiation of EGF trafficking, cells were fixed and stained with antibodies against EEA1, CD63, or LAMP1 by using anti-FITC detection (green). Cells were imaged by confocal microscopy. (B) The extent of overlap between the indicated endosomal markers and EGF-TR was determined using MetaMorph software as described in Materials and Methods. For each condition, 30–50 cells were imaged from three independent experiments. The percentage of EGF-TR that colocalizes with marked endosomes.