|
|
|
|
|
|
|
|
|||||
| Home Help Feedback Subscriptions Archive Search Table of Contents | |||||
|
Fig. 3. Single-molecule imaging of the transient translocation of Crac-GFP to membranes upon stimulation with cAMP. (A) Sequential images of a cell expressing Crac-GFP observed under TIR-FM (see supplementary material Movie 1). The cell was stimulated at 0 seconds with cAMP. (B) Temporal changes in the number of Crac-GFP spots on membrane. After stimulation with cAMP, the number of the Crac-GFP spots increased transiently 
10 second. (C) Dissociation of Crac-GFP from membranes of cells stimulated with cAMP. Lifetimes of Crac-GFP between initial observation and disappearance on membrane were measured and plotted as a cumulative curve, giving rise to the dissociation curve. The data can be fitted to single exponential curve with a constant (
) of 120 milliseconds, indicating rapid exchanges of individual Crac molecules during Crac recruitments. Dissociation curve of Crac-GFP in living cells with =120 milliseconds (
) and photobleaching curve of Crac-GFP absorbed on the surface of coverslip with =3800 milliseconds (
).
|
