LSCM investigations of pardaxin disruption of GUVs. For each lipid, we show the time course (in minutes) of the fluorescence from a water-soluble fluorophore (Alexa 488, encapsulated within the vesicle interior) in the top row and a membrane-bound dye (DiI) in the bottom row after the addition of 5 mM pardaxin to GUV composed of either DOPC or DOPG/DOPC (20:80). For DOPC GUVs we start to observe a decrease in the water-soluble fluorophore fluorescence after ∼29 min, reaching a minimum after 45 min, whereas the DiI fluorescence is unchanged. For the DOPG/DOPC (20:80) GUVs, we observe a decrease in the GUV size after roughly 40 min, leading to vesicle disruption within 5 min indicated by the disappearance of both fluorophores. Note that there is a slight inhomogeneity in the dye distribution in the DOPC vesicle. However, the same type of behavior, indicative of pore-formation, is observed for a large range of different vesicle sizes for DOPC irrespective of the dye distribution.