Inhibition of myosin-Vb after loading with transferrin does not prevent transit from perinuclear recycling endosomes. HeLa cells transiently expressing sensitized myosin-Vb were loaded with Alexa 546-transferrin, washed, incubated in growth medium for 30 min, and imaged for myosin-Vb and transferrin. (A) Diagram depicting predicted results; the sensitized mutant myosin-Vb is shown in red and PE-ADP is shown as a green circle. (B) Inhibition of accumulation of transferrin (red) added after myosin-Vb inhibition by microinjection of PE-ADP2. Injected cells have blue nuclei. (C, D, E, F, G, H) The cell expressing sensitized myosin-Vb (center, panel C) was immediately injected with PE-ADP and the same field was imaged 30 min later (F, G, H). Panels C and D are overlaid in panel E, and panels F and G are overlaid in panel H. Bar, 15 μm.
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