Fig. 4: Triple-layer inverted emulsion production of phase-separated GUVs.
From: Engineering synthetic cells with intramembrane domains possessing distinct bilayer asymmetries
a Phase diagram of three-component lipid bilayer showing two-phase and single-phase regions produced from ternary mixtures with varying concentrations of cholesterol. DOPC:Sphingomyelin were at a fixed 1 : 1 molar ratio, and in additional GUVs are labeled with 0.5 mol% DiD which partitions into the Ld phase. Black dots represent the concentrations which resulted in co-existing phases (a vesicle population with more than 50% of the GUVs exhibiting domains), and red dots represent the concentrations that resulted in a single-phase (less than 50% with visible domains). b Comparing the percentage of phase separation amongst the GUVs produced with different cholesterol concentrations (DOPC:SPM – 1 : 1) between the triple-layer method and the electroformation (EF) method. c Confocal images (DiD signal) of phase-separated GUVs produced using the triple-layer method with cholesterol percentages of 10%, 20%, 30%, 40%, 50%, 60%, 70% and 80%. n ≥ 70, data is collected from three or more independent experiments. Scale bars correspond to 20 µm.
