Figure 7.

EhVps20, EhVps32 and EhVps24 are required to produce ILVs in GUVs and EhVps2 apparently modulates their size. (A) GUVs with the same composition as in Figure 6 and each recombinant protein were added and incubated at RT with 10 min intervals. As a negative control, GUVs were incubated for the same times replacing proteins with buffer. (B,C) Experiments where some protein was omitted or substituted as indicated at right. Scale bar: 10 μm. (D) Quantification of the number of GUVs containing ILVs formed for subsequent addition of the proteins indicated below. In the bottom of each bar images of ILVs formation after each protein addition are shown. (E) GUVs prepared with DOPC alone or a mixture of different negatively charged lipids (negative molar ratio of 10%) were incubated with the four ESCRT-III proteins. Bars represent the mean and standard error of three independent experiments.