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. 2006 Jul 31;174(3):459–471. doi: 10.1083/jcb.200508170

Figure 3.

Figure 3.

TeNT HC enters clathrin-coated structures in MNs. (A and B) MNs were microinjected with a plasmid encoding GFP-CLC. After overnight expression, cells were incubated with 30 nM Alexa Fluor 555–TeNT HC, washed, and imaged. (A) GFP-CLC and TeNT HC dual-positive structures are visible in an axon (arrows). (B) Confocal section of the bottom plane of a MN soma. A partial overlap between GFP-CLC–positive and TeNT HC–positive structures is observed (inset). (C) MNs were incubated with HRP–TeNT HC for 45 min on ice, and then chased for 45 min at 12 (a and b) or 18°C (c). Cells were fixed and incubated with DAB/H2O2 to label HRP–TeNT HC. The electron-dense DAB reaction product was often associated with coated pits and budding vesicles, which were visible at higher magnification (b' and c'). Bars: (A) 2 μm; (B) 5 μm; (C) 0.2 μm.