Figure 10

Morphology changes lead to visible membrane permeability only at a Rh2/lipid ratio of 6. GUVs composed of eSM:ePC (A), ePC:Chol (B) or eSM:ePC:Chol (C) were labeled with Rhod-PE (red) and diluted in 0.2 M glucose supplemented with 20 μM calcein (green). GUVs were incubated with either 1% DMSO (line 1) or with Rh2 at a Rh2/lipid ratio of 3 after 1 or 10 min (lines 2 and 3, respectively), or of 6 for 5 min (line 4) at 22 °C. The difference of green fluorescence intensities between outside and inside vesicles was measured and indicated in the merge panel. GUVs were visualized by confocal microscope with Axio Observer 1 inverted microscope equipped with a spinning disk microscope. Scale bars, 20 µm. One-way ANOVA with Bonferroni’s multiple comparison. ***p < 0.001. Images are representative of two independent experiments and at least 15 GUVs were analyzed per experiment.