Fig. 3: Cryo-EM shows that asymmetrical glycerophospholipids form thinner membranes and support phase separation with either ergosterol or diplopterol.
a Representative cryo-EM images of single component LUVs made from the monounsaturated symmetrical lipid POPC (top panel) and the asymmetrical saturated SDPC (bottom panel). Right, magnified areas of respective images. b Representative normalized intensity profiles for POPC and SDPC liposomes. The trough-to-trough distance DTT is shown by arrows within dotted lines. c Representative cryo-EM images of two-component liposomes made from either POPC or SDPC glycerophospholipids in combination with 30 mol% ergosterol or diplopterol. The gel-like membranes composed of the symmetrical saturated lipid DPPC with ergosterol is also included. Magnified areas are shown at the bottom of respective images. d Representative cryo-EM images of three-component liposomes made either from 40 mol% POPC or SDPC glycerophospholipids in combination with 40 mol% gel-forming DPPC and 20 mol% ergosterol or diplopterol. Magnified areas are shown at the bottom of respective images. Note the phases of different membrane thickness in the ternary SDPC-DPPC-based membranes. e Representative normalized intensity profiles for thick and thin regions in phase-separated liposomes made from SDPC in the presence of DPPC and ergosterol or diplopterol. The trough-to-trough distance DTT is indicated by arrows. f The violin plot shows DTT values for single, two- and three-component LUVs assembled with POPC or SDPC (average ± S.D. n = 20 liposomes, 10 measurements for each liposome, one biological repeat). Medians are indicated. p values were obtained from two-tailed unpaired parametric t-tests. (a, c, d) Scale bars represent 20 nm. Source data are provided as a Source Data file.
