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. Author manuscript; available in PMC: 2020 Dec 16.
Published in final edited form as: Angew Chem Int Ed Engl. 2019 Oct 31;58(51):18683–18690. doi: 10.1002/anie.201911544

Figure 2.

Figure 2.

Phase segregation enhances DNA-mediated fusion. Maximum intensity projection of z-stacked fluorescence micrographs of GUVs containing DSPC: DOPC: DOPE: Chol at a molar ratio of (a) 0:3:1:1 and (b) 2:1:1:1 and with 0.1 mol% Rhod-PE, which localizes to the liquid-disordered phase. Scale bars are 5 μm. (c)Total lipid mixing for homogenous (0 DSPC: 3 DOPC: 1 DOPE: 1 Chol) and phase segregated vesicles (2 DSPC: 1 DOPC: 1 DOPE: 1 Chol). cDNA total lipid mixing for both lipid mixtures was significantly different from respective controls and from one another (p < 0.0001, Two-Way ANOVA Tukey Test). (d) Content mixing assay for homogenous and phase segregated vesicles reports increases in calcein fluorescence that accompany vesicle fusion. Values are normalized to the percent increase in calcein fluorescence observed upon mixing of homogenous vesicles functionalized with cDNA tethers. ncDNA and cDNA were not significantly different for 0 DSPC: 3 DOPC: 1 DOPE : 1 Chol (p > 0.05, Sidak’s Two-Way ANOVA), cDNA of 2 DSPC: 1 DOPC: 1 DOPE: 1 Chol was significantly different than all other samples (Sidak’s Two-Way ANOVA). Lipid and content mixing assays were carried out at 25ºC at pH 7.3 in PBS. Error bars represent standard error of the mean (S.E.M), n=3.