Movie 4.

GFP is not recruited to the membrane during toxin challenge. GFP (green) transfected HeLa cells were challenged with sublytic (A) SLO (250 HU/mL), (B) ILY (250 HU/mL), (C) aerolysin (Aero) (62 HU/mL), or (D) mass equivalent to aerolysin of aerolysin^Y221G (AeroY221G) in imaging buffer including 2 μg/mL TO-PRO3 (blue) and imaged at 37°C by live cell confocal imaging at 1-3.5 frame/second. Triton-X-100 was added just prior to the end of imaging as a positive control for cell permeabilization. Images were bleach-corrected by histogram matching. Time displays min post toxin challenge. Scale bar = 5 μm.