Fig. 1: Encapsulation of bundled actin in giant unilamellar vesicles.
From: Reconstitution of contractile actomyosin rings in vesicles
a Schematic depiction of the vesicle generation process. The aqueous protein solution is injected into a rotating chamber through a glass capillary. Droplets form at the capillary tip in the oil phase, which contains lipids. The droplets then pass through a water–oil interphase lined with a second lipid monolayer, forming the giant unilamellar vesicles (GUVs). b Field of view image (Z-projections of confocal stacks) with many cytoskeletal vesicles. Actin in green. See Supplementary Movie 1 for 3D effect. c Comparison of cytoskeletal vesicles with actin bundled by four different types of bundling proteins. We used 2 µM actin in all cases, but due to differences in bundling activity, different concentrations of bundling protein: 0.3 µM fascin, 0.9 µM VASP, 1 µM α-actinin, 2 µM talin, and 2 µM vinculin. d Automated tracing of bundles by analysis script. Confocal z-stacks are converted into a three-dimensional “skeleton” model. Supplementary Movie 2 shows a 3D view of both representations of these vesicles.
