Fig. 1.

ENTH deforms membranes in a PS-dependent manner. a SDS-PAGE of co-sedimentation shows that ENTH domain binding to LUVs only depends on the presence PI(4,5)P₂. LUVs consisted of polar brain lipid extract (Folch) in the absence and presence of PIP2 as indicated (Folch: PI(4,5)P₂ (95: 5 mol%)). b ENTH domain induced membrane deformation upon LUV binding as analyzed by electron microscopy. Lipid composition as in (a). c Dynamic light scattering spectra of LUVs shown in (b) showed PIP2-dependent alterations DLS data were acquired by 3 times 3 independent measurements, each one consisting of 21 repetitions for each lipid composition. Error bars represent the standard deviation. d SDS-PAGE of co-sedimentation of ENTH domain with LUVs of indicated lipid composition LUVs containing PS were composed of PC, PE, PS and PI(4,5)P₂ (20% PS—45: 30: 20: 5 mol%; 5% PS—60: 30: 5: 5 mol%). LUVs without PS were composed of PC, PE and PI(4,5)P₂ (0% PS—65: 30: 5 mol%) and LUVs containing PI were composed of PC, PE, PI, PI(4,5)P₂ (20% PI—45: 30: 20: 5 mol%). e Statistical evaluation of EM micrographs shown in (f). Error bars represent the standard error of the mean (SEM). f Electron microscopy of negatively stained LUVs with indicated increasing PS or PI concentrations. LUVs were composed with similar lipid compositions as in (d). At least 250 membrane structures were counted for each condition and the error bars represent the standard deviation. g Dynamic light scattering spectra of LUVs shown in (e). DLS data was acquired by 3 independent measurements, each one consisting of 21 repetitions for each lipid composition. Each measuring circle was repeated 3 times