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Lysenin life cycle start with a soluble monomer that binds to membrane lipid sphingomyelin. After an oligomerization process Lysenin nonamers, known as prepores, suffer endocitosis. In the late endosome the acidic conditions induce the oligomer insertion, resulting in the so-call pore state. High-Speed Atomic Force Microscopy allow to study Lysenin life cycle, highlighting the crucial role of crowdedness in it.