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. 2011 Oct 3;195(1):71–86. doi: 10.1083/jcb.201011112

Figure 4.

Figure 4.

FIP5 is required for the scission of endocytic carriers at recycling endosomes. (A) Control (left) and GFP-FIP5-F1–expressing (right) HeLa cells were incubated with Tf-HRP for 45 min at 37°C. Cells were then fixed and prepared for ultrastructural EM analysis. Electron-dense dark precipitate indicates DAB reaction product and highlights the localization of Tf-HRP. (B) GFP-FIP5-F1–expressing HeLa cells were incubated with Tf-HRP for 45 min at 37°C. The DAB reaction was performed before permeabilization, antibody incubation, and fixation. Electron-dense dark precipitate indicates the DAB reaction product and highlights the localization of Tf-HRP, whereas 10 nm gold particles indicate the localization of GFP-FIP5-F1 (arrows). (C–F) Higher-magnification images from HeLa cells expressing GFP-FIP5-F1 and loaded with Tf-HRP. Asterisks and arrows in C and D point to the “bead” that lacks Tf-HRP, whereas arrowheads track nearby microtubules. Bars: (A) 5 µm; (B–F) 200 nm.