Fig. 1: GFP photobleaching analysis via SiMPull-POP reports on EphA2 oligomerization in a native-like membrane environment.
From: Cholesterol inhibits assembly and oncogenic activation of the EphA2 receptor
a Schematic representation of sample preparation and workflow for SiMPull-POP. 1- Membrane fractions containing EphA2-GFP were solubilized with the amphipathic copolymer DIBMA to generate DIBMALPs. 2- EphA2-GFP DIBMALPs were immobilized on a functionalized microscope slide displaying an EphA2 antibody. 3- DIBMALPs devoid of EphA2-GFP are washed away before imaging. b Representative single-molecule TIRF image in the presence (left) and absence (right) of EphA2 antibody (scale bar = 5μm). Each blue spot represents a DIBMALP containing EphA2-GFP. c Representative GFP photobleaching traces showing a stepwise decrease in GFP intensity over time; arrows represent individual photobleaching events. Photobleaching steps are used to infer EphA2 oligomerization status.
