Movie 1.

Imaging-based protease protection assay with full-length Hobbit-GFP. Live-cell timelapse imaging of cytosolic mTagBFP2 (cyan), full-length Hobbit-GFP (green), and KDEL-RFP (red) in an imaging-based protease protection assay. The selective detergent digitonin, which permeabilizes the plasma membrane but not intracellular membranes, was added at timestamp 28.049430 s, and proteinase K was added at timestamp 2 min 4.664132 s. Still images from this time-lapse are pictured in Fig. 4C.