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a Phase diagram of three-component lipid bilayer showing two-phase and single-phase regions produced from ternary mixtures with varying concentrations of cholesterol. <t>DOPC:Sphingomyelin</t> 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 <t>(DOPC:SPM</t> – 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.
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a Phase diagram of three-component lipid bilayer showing two-phase and single-phase regions produced from ternary mixtures with varying concentrations of cholesterol. <t>DOPC:Sphingomyelin</t> 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 <t>(DOPC:SPM</t> – 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.
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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.

Journal: Nature Communications

Article Title: Engineering synthetic cells with intramembrane domains possessing distinct bilayer asymmetries

doi: 10.1038/s41467-026-68997-x

Figure Lengend Snippet: 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.

Article Snippet: 1-palmitoyl-2-oleoyl-glycero-3-phosphocholine (POPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), Sphingomyelin (SPM), Cholesterol (Chol), 1,2-dioleoyl-sn-glycero-3-[(N-(5-amino-1-carboxypentyl)iminodiacetic acid)succinyl] (nickel salt) (DGS-Ni-NTA), GM1 Ganglioside (Brain, Ovine-Sodium Salt) (GM1), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(cap biotinyl) (sodium salt) (biotinyl Cap PE), and L-α-Phosphatidylethanolamine-N-(7-nitro-2-1,3-benzoxadiazol-4-yl) (Ammonium Salt) (NBD-DOPE) are purchased from Avanti polar lipids.

Techniques: Produced, Labeling