Supplemental Material

This article contains the following supporting material:

• Supplemental Figures 1-8.pdf
• Movie 1 - CT traffics in vesicles and tubules.
  BSC1 cells were incubated with 20nM CT(E112D) at 37°C for 45min, and confocal images were captured every ~3sec for a total of 5min.
  
• Movie 2 - Vesicles and tubules carrying CT move along microtubules.
  BSC1 cells transiently expressing tubulin-EGFP for 20h, were incubated with 20nM CT(E112D) at 37°C for 45min, and images were captured every 2sec for a total of ~2min. Last section of movie shows a duplicate of only CT traffic to aid visualization of vesicles and tubules.
  
• Movie 3 - Caveolin structures mediate CT internalization.
  BSC1 cells transiently expressing caveolin1-EGFP for 24h were placed inside a chamber for live-cell imaging and stabilized at 37°C with HMEMB medium. Time-lapse recording started immediately before addition of 20nM CT(E112D), and images were captured every 30sec for a total of ~25min.
  
• Movie 4a - CT-induced morphological change.
  Y-1 cells placed inside a chamber for live-cell imaging and stabilized at 37°C with HMEMB medium. Timelapse recording started immediately before addition of wild type CT (movie 4a) or 10μM forskolin (movie 4b), and phase-contrast images were captured every 30sec for a total of ~70-80min. The morphological change induced by CT had a much longer lag-phase (~35min) compared to forskolin (~2min), consistent with the need for the toxin to traffic from the plasma membrane to the ER, where CT is retro-translocated into the cytosol and then induces toxicity.
  
• Movie 4b

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