Movie 3.

Intrinsic repair drives shedding of A6⁺ microvesicle following aerolysin challenge. HeLa cells transfected with A6-YFP (green) were challenged with sublytic (A) SLO (250 HU/mL), (B) pro-aerolysin (62 HU/mL), (C) aerolysin (aero) (62 HU/mL), or (D) mass equivalent of aerolysin^Y221G (Aero^Y221G) to aerolysin 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 after toxin challenge. A6-YFP recruitment to the sites of plasma membrane damage are shown by arrowheads. Scale bar = 10 μm.