Fig. 4: In vitro pore formation assay for determining inhibitor effectiveness.
A A schematic of the assay showing how pore formation is used as an indicator of toxin maturation. His6-CylA-27-412 cleaves mCylLS and mCylLL producing CylLS” and CylLL”, which cooperatively form pores in pyranine-encapsulated DOPC liposomes at nanomolar concentrations. Pore formation dissipates a pH gradient between the interior of the liposome and the surrounding buffer increasing pyranine fluorescence. B Representative example of changes in membrane permeabilization activity with decreasing concentration of 1. C Representative plot of the permeabilization activity as a function of the concentration of 1 fit to a dose-response curve. Fitting was done using Origin Pro 2024 yielding EC50 values (see Supplementary Fig. 3 for additional data for compounds 2-10). The data points from two replicates are shown (n = 2). Source data are provided as a Source Data file.
