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. 2009 Dec 7;30(3):781–792. doi: 10.1128/MCB.00330-09

FIG. 3.

FIG. 3.

Tf addition stimulates an increase in vesicle budding from the PM. (a) A low-magnification TIRF microscopy image of living Clone 9 cells expressing Dyn2-GFP. A series of higher-magnification images of the boxed region in panel a are shown in panels b to d. Cells in the absence of Tf showed little, if any, Dyn2-GFP-based vesicle budding over the 10- to 20-min observation time (b). In contrast, addition of Tf (5 μg/ml) induced the formation of Dyn2 vesicles from the PM that continued over a 20-min time period (c and d). Statistical counts of this observed Dyn2-GFP vesicle formation within 10-μm by 10-μm boxes of 5 different cells viewed 10 min before and 10 min following Tf addition showed a >40% increase in Dyn2 vesicle formation subsequent to ligand addition. Bar = 10 μm. (e) IP of the TfR1 from MEFs showing an increase in associated Dyn2 and Cort minutes following Tf addition. (f and g) Densitometric quantitation of 3 independent experiments similar to that shown in panel e comparing the ratio of Dyn2 and Cort to TfR1. Results represent the average ± SE of 3 independent experiments.