Xenbase Image ID: 122531

Figure 3. Live-cell imaging of tagged CVAK104 constructs.A and B) CVAK104 was tagged with GFP at the C-terminus and transiently expressed in HeLa cells. A) In control cells, CVAK104-GFP is present in small and highly mobile compartments. B) In clathrin-depleted cells, CVAK104-GFP is more cytosolic but also labels largely immobile vesicular compartments as well as mobile tubular structures (white arrows). C and D) The N-terminal kinase-like domain of CVAK104 was tagged with YFP and transiently expressed in HeLa cells. C) In control cells, the construct localizes to mobile tubular and vesicular structures. D) In clathrin-depleted cells, the construct localizes mainly to tubular mobile structures. E) CVAK104-GFP was transiently expressed in HeLa cells. Cells were allowed to endocytose pre-bound Alexa Fluor-labelled transferrin, and imaged either ∼1, ∼10 or ∼60 minutes after beginning of uptake by spinning-disk confocal microscopy. Frequent colocalization (white arrows) can be observed at early, but not at late stages of the experiment. Time-points 1 and 10 minutes show images of the same cell, taken from a continuous movie (Video S5). Representative still images are shown here; the corresponding movies are available as Videos S1–S6 (online supplemental material). Scale bars: 10 μm. Image published in: Borner GH et al. (2007) Image downloaded from an Open Access article in PubMed Central. © 2007 The Authors Journal compilation © 2007 Blackwell Publishing Ltd Larger Image Printer Friendly View

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