A) Fluorescence anisotropy vs. temperature of synthetic vesicles composed of GlcCer from the wild-type (left), Δsmt1 (middle), and Δsld8 strains (right). A sigmoid function was fit to the anisotropy profiles and the chain melting temperature (Tm) was defined as the inflection point of that sigmoid. Error bars represent the standard deviation from three experiments for the wildtype and two experiments for the mutant strains. B) Images of giant unilamellar vesicles (GUVs), synthesized from GlcCer, POPC, and ergosterol (Erg), in a 1:1:1 molar ratio. GlcCer purified from the wild-type strain (left), Δsmt1 strain (middle), and Δsld8 strain (right) was mixed with equimolar concentrations of POPC and Erg. Rhodamine-DOPE (0.01 mol%), localizing in disordered lipid phases, was used as the fluorescence probe.