Fig. 5: Asymmetric phase-separated synthetic cells with sphingomyelin using the triple-layer inverted-emulsion method.
From: Engineering synthetic cells with intramembrane domains possessing distinct bilayer asymmetries
a Schematic representation of the asymmetric phase separated GUVs with biotinylated lipids (light green) binding streptavidin (green) in the inner leaflet of the Ld domain (black region) only, and GM1 (blue) binding CT-B Al-594 (red) in the outer leaflet of the Lo domain (pink region) only. b Confocal images of asymmetric phase-separated GUVs (50 mol% Chol, DOPC:SPM – 1:1, 1 mol% GM1 in the outer leaflet and 50 mol% Chol and DOPC:SPM – 1:1, 0.2 mol% biotinyl cap PE in the inner leaflet), showing streptavidin (green) binding to the liquid disordered domain, and CT-B Al-594 (red) binding to the Lo domain. Scale bars correspond to 20 µm. c Time-dependent curvature dynamics induced within the Lo domain of the asymmetric phase-separated vesicles after the addition of 20 nM CT-B Al-594. Scale bars correspond to 20 µm. d Time series of osmotic deflation induced budding-off of the curved CT-B AL-594 Lo domain (in red). Scale bar corresponds to 10 µm. e Time-series of osmotic deflation induced division of an Lo daughter vesicle from the Ld domain. Data is collected from three or more independent experiments. Scale bar corresponds to 20 µm.
