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International Journal of Cell Biology
Volume 2013, Article ID 560421, 18 pages
Research Article

Early Delivery of Misfolded PrP from ER to Lysosomes by Autophagy

1Departments of Neurology, MC2030, The University of Chicago Pritzker School of Medicine, 5841 S. Maryland Avenue, Chicago, IL 60637, USA
2Departments of Neurobiology, The University of Chicago Pritzker School of Medicine, Chicago, IL 60637, USA

Received 12 May 2013; Accepted 20 September 2013

Academic Editor: Roberto Chiesa

Copyright © 2013 Constanza J. Cortes et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Supplementary Material

Video Supplement S1. Time-lapse fluorescence microscopy of Mut-PrP. Maximum intensity projection of time-lapsed confocal z-stack sequence for a COS-7 cell observed every 10 min from the first detection of Mut-PrP::GFP (~ 5 h) for 2 h and displayed at 3 fps. Cells were located using the EM-CCD, as the expression level was poorly visible to the eye. Note that the earliest detection of Mut-PrP is in the form of puncta, suggesting aggregates of PrP. This is supported by the finding that these puncta increase in intensity and size over the observation period. Also note the perinuclear accumulation of puncta. This movie corresponds to Figure 3A.

Video Supplement S2. Time-lapse fluorescence microscopy of Mut-PrP with LysoTracker Red. Images are as described in Video Supplement 1. Acidic vesicles are labeled with LysoTracker Red. Images were collected every 10 min over a 140 min observation period, beginning at T=7 h (420 min) and displayed at 3 fps. Colocalized pixels are rendered solid yellow to aid visualization. This corresponds to Figure 3B.

Video Supplement S3. Time-lapse fluorescence microscopy of Wt-PrP with LysoTracker Red. At T=3 h, cells were imaged every 10 min for 950 min. Images are maximum intensity projections using only the central optical slices of the cell (to visualize internal details), created by trimming the z-stack data using ImageJ software. Image display rate is 3 fps. This corresponds to Figure 3C.

  1. Supplementary Materials