Figure 3 : CD59 releases an ILY late-prepore independent of cholesterol.

From: Disentangling the roles of cholesterol and CD59 in intermedilysin pore formation

Figure 3

(a) Schematic illustrating the liposome flotation assay. Liposomes decorated with cytoCD59 were incubated with ILY and subjected to density centrifugation. Monomeric and oligomeric forms of the toxin in each fraction were separated by SDS-AGE and detected by western blot analysis. Samples containing cholesterol-rich liposomes and the ILYTI variant are shown in (b), while those in (c) contain wildtype ILY prepores formed on liposomes lacking cholesterol. Fractions from a single gradient span two gels (left and right). Directionality of fractionation is indicated by Top and Bottom above gels. The first lane in each gel contains molecular weight markers. (d) Cryo-electron micrograph of the sample analyzed in panel c showing ILY oligomeric prepores on liposomes (white arrows) and those released from the membrane (red arrows). Scale bar, 50 nm.