Fig 3: Lipid segregation in cholesterol and CHIM containing membranes.

a Top view of a DPPC:DLiPC:sterol bilayer. DPPC is shown in green, DLiPC in gray, CHOL in orange, and CHIM in blue. The top row shows snapshots of the initial configuration while the bottom row shows snapshots of the final configuration after a simulation of at least 3 µs. b Top: Average ratio of DPPC neighbors over the total number of PL neighbors around DPPC averaged over the last 25% of the simulation. Bottom: Average ratio of DPPC neighbors over total PL neighbors around CHOL or CHIM. The dashed lines at 0.9 indicate complete segregation and at 0.4 complete mixing. c GUVs composed of POPS:DLiPC:DPPC:CHIM-L (14.29:42.85:28.57:14.29), stained with FAST DiI and clicked with AF488. Lipid segregation occurs at RT, as revealed by FAST DiI labeling of the Ld phase. Note that CHIM-L (labeled by the click reaction with AF488) shows a tendency to partition into Lo, as also seen in the line plot on the right. d DCA-treated GPMVs derived from HUVEC labeled with FAST DiI and fluorescently labeled CHIM-L (AF488). Note that CHIM partitions into the more ordered phase (Lo) negative for FAST DiI. The corresponding line plot (fluorescence intensity) for the vesicle is displayed on the right.