Fig. 2: Asymmetrical glycerophospholipids support membrane phase properties both in the presence of ergosterol and diplopterol. | Nature Communications

Fig. 2: Asymmetrical glycerophospholipids support membrane phase properties both in the presence of ergosterol and diplopterol.

From: Horizontal acquisition of prokaryotic hopanoid biosynthesis reorganizes membrane physiology driving lifestyle innovation in a eukaryote

Fig. 2

a Chemical structures of glycerophospholipids used in the study. b Estimation of membrane order from spectral GP imaging of two-component GUVs made with POPC or SDPC with either ergosterol or diplopterol. Control values for gel-like DPPC/Erg GUVs are shown for reference. The values represent average ± S.D. (n = 15 GUVs, one biological repeat), estimated using Eq. (2). The top panel shows representative spinning disk confocal images (mid-plane) of three-component GUVs assembled with equimolar amounts of symmetrical unsaturated glycerophospholipids POPC or DOPC and the gel-like DPPC (c), or asymmetrical saturated glycerophospholipid SDPC (d) with either 20 mol% ergosterol or diplopterol. GUVs were labeled with FAST DiI. Scale bars represent 2 μm. Dot plots depict the quantification of area of liquid-disordered (Ld) regions upon phase separation (average ± S.D., with the following sample sizes: POPC DPPC Dip (n = 20 GUVs), DOPC DPPC Dip (n = 20 GUVs), POPC DPPC Erg (n = 71 GUVs), DOPC DPPC Erg (n = 66 GUVs) SDPC POPC Erg (n = 20), SDPC POPC Dip (n = 20), SDPC DPPC Erg (n = 58) and SDPC DPPC Dip (n = 60), from two biological repeats). e–g Quantification of order in two phases of three-component GUVs made with the following lipids (40/40/20 mol%): POPC, DPPC, and ergosterol (e), SDPC, DPPC and ergosterol (f), and SDPC, DPPC and diplopterol (g). Lo represents the liquid-ordered regions, and Ld represents liquid-disordered regions. GP values were calculated using Eq. (2). Average ± S.D. are shown (n = 12 GUVs, one biological repeat). h Representative single-plane pseudo-colored generalized polarization (GP) images. Color bar indicates the range of GP values where blues show low membrane order and reds show high membrane order. Areas quantified are outlined in white. Zoomed regions show plasma membrane and nuclear membrane areas used to quantify GP values. Scale bar, 5 µm. i A plot representing individual GP values quantified at the plasma membrane regions at cell tips and at the nuclear membrane in cells of indicated genotypes. Average ± S.D. are shown (n = 25 cells from three biological repeats). (b–g, i) p values were obtained by two-tailed unpaired parametric t-tests. Source data are provided as a Source Data file.

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