Figure 6.
Figure 6. Cytohesin-1 localization is defined by G272. (A) HeLa cells stably expressing EGFP-tagged diglycine CYTH1 were either not treated (−, unfilled) or treated (+, filled) with HGF for the indicated time points, permeabilized with ice-cold 0.05% saponin in Pipes buffer, and imaged by confocal microscopy. (B) HeLa cells stably expressing EGFP-tagged variants of CYTH1 were either not treated (−, unfilled) or treated (+, filled) with HGF for 15 min, permeabilized with ice-cold 0.05% saponin in Pipes buffer, and imaged by confocal microscopy. (C and D) Cells were prepared as in A, except they were pretreated with DMSO, 0.2 µM Wortmannin, or 20 µM LY294002 for 30 min before HGF stimulation. (E) HeLa cells stably expressing triglcyine EGFP-CYTH1 were pretreated with 10 µM DMSO or ionomycin for 10 min, permeabilized with ice-cold 0.05% saponin in Pipes buffer, and imaged by confocal microscopy. Scale bars, 20 µm.

Cytohesin-1 localization is defined by G272. (A) HeLa cells stably expressing EGFP-tagged diglycine CYTH1 were either not treated (−, unfilled) or treated (+, filled) with HGF for the indicated time points, permeabilized with ice-cold 0.05% saponin in Pipes buffer, and imaged by confocal microscopy. (B) HeLa cells stably expressing EGFP-tagged variants of CYTH1 were either not treated (−, unfilled) or treated (+, filled) with HGF for 15 min, permeabilized with ice-cold 0.05% saponin in Pipes buffer, and imaged by confocal microscopy. (C and D) Cells were prepared as in A, except they were pretreated with DMSO, 0.2 µM Wortmannin, or 20 µM LY294002 for 30 min before HGF stimulation. (E) HeLa cells stably expressing triglcyine EGFP-CYTH1 were pretreated with 10 µM DMSO or ionomycin for 10 min, permeabilized with ice-cold 0.05% saponin in Pipes buffer, and imaged by confocal microscopy. Scale bars, 20 µm.

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