Supplementary material for Cutler et al. (2000) Proc. Natl. Acad. Sci. USA 97(7), 3718-3723.
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Shown is a time series acquired from a hypocotyl epidermal cell of transgenic line C2 expressing an out-of-frame GFP fusion protein. The time series shows several torus-shaped organelles. There appear to exist two populations: doughnut-shaped organelles and more motile tubular forms of the same organelle. Examples of the doughnut-shaped structures converting into motile tubular forms can be seen throughout the movie. Images frames were acquired at 0.75-second intervals and are shown in the movie at 10 frames per second.
Shown is a time series acquired from hypocotyl epidermal cells in a transgenic line expressing a GFP::delta TIP fusion protein. The time series demonstrates the remarkably dynamic nature of the vacuolar membrane system. Image frames were acquired at 4.5-second intervals and are shown in the movie at 10 frames per second.
Shown is a time series acquired from leaf petiole epidermal cells of EGAD line Q10, which expresses an in-frame fusion protein to a glycine-rich protein of unknown function. The marker highlights dynamic bubble-like structures and faintly marks the ER membrane. The bubbles seem to be ER-associated in that their movement is highly correlated with the background ER fluorescence. Image frames were acquired at 3-second intervals and are shown in the movie at 10 frames per second.
Time series at one confocal plane of several hypocotyl epidermal cells of transgenic line Q4, which expresses an in-frame GFP fusion to a novel protein. The marker is localized to what is likely the ER membrane, based on both colocalization with BODIPY FL Ceramide and similarity in appearance to KDEL-tagged GFP (made by Jim Haseloff, Medical Research Council, Cambridge, U.K.). The visible network appears to be composed of elements of varying stability; a more stable peripheral network and rapidly remodeled components likely associated with cytoplasmic strands. Note the mobile cigar-shaped structures at the right of the image. Similar structures have been seen in plants expressing KDEL-tagged GFP. Although KDEL-GFP appears to label the lumen of these structures, Q4-GFP localizes to their surface. The identity of these distinctive structures is not known. Images in the series were acquired at 2-second intervals and are shown in the movie at 10 frames per second.
Shown is a time series taken from a hypocotyl cell of EGAD line F2, which expresses an out-of-frame fusion protein. The marker illuminates the outside of a spherical oganelle that contains a large asymetric dot. The identity of this organelle is unknown. Several Q-balls can be seen moving toward the nucleus in the time series. Image frames were acquired at 4.5-second intervals and are shown in the movie at 10 frames per second.