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Fig. 1. Distinct elements of cytoskeleton are essential for sequestration of ß₂AR induced by stimulation of cells with insulin (+Ins) or isoproterenol (+Iso). A431 cells stably expressing ß₂AR tagged with enhanced GFP were used to monitor receptor internalization (by confocal microscopy) following stimulation with ß-adrenergic agonist (10 µM isoproterenol) or 100 nM insulin. Control cells, without any pretreatment or cells treated with cytoskeletal disrupting drugs nocodazole (10 µM) or latrunculin (1 µM) were subsequently stimulated with either insulin (100 nM, +Ins) or isoproterenol (10 µM, +Iso) for 30 minutes. After the fixation and embedding in SlowFade, cells were analyzed by confocal microscopy. In the control experiment (top panel), receptor relocated from cell periphery (area of plasma membrane, marked with yellow arrows) to nuclear proximity (white arrowheads). In the presence of nocodazole (+Noc.), stimulation with insulin (+Ins) induced sequestration of receptor from plasma membrane, whereas in cells stimulated with isoproterenol (+Iso), receptor remained on cell periphery (yellow arrows). Pretreatment of cells with latrunculin (+Latr.) suspended ß₂AR relocation in response to stimulation with insulin. Bars, 10 µm.

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