JCS01167 Supplemental Data

Files in this Data Supplement:

• Supplemental Data (PDF)
• Movie 1 -

  Movie 1. Three-D image of live Xenopus retina. This 3-D image was constructed from the CLSM scans that were used to generate Fig. 3 of the manuscript. In general in the experiments, we set the excitation beam to be the maximum intensity that did not cause saturation of the avalanched (photon-counting) photodiodes. The stack of scans were imported as 8-bit images into Volview^™, and the movie generated with its macro-facility. Because some of the rods have much lower expression than others, a linear gradient of display was used for the lowest 50% of the intensities in the images, allowing the lower expressing rods to be seen more clearly. For quantitative processing as shown in Fig. 5, the raw scan values (counts ms^–1) were always used.

• Movie 2 -

  Movie 2. Single Xenopus rod expressing EGFP. This rod cell is the one that is identified in Fig. 3B with a white arrow and is also featured in panel A of Fig. 4. Only the voxels in the top 5% of the fluorescence intensity distribution have been shown in green. The lowest 5% are shown in red, highlighting primarily the boundary of the cell. All other voxels have been ‘turned off’. A linear opacity gradient has been used. This image should be compared with that in Movie 3.

• Movie 3 -

  Movie 3. Dark-adapted Xenopus rod expressing the arrestin-EGFP fusion protein Arr-EGFP rod. This is the same rod as illustrated in Fig. 7A,D and Fig. 8A,D. To highlight the region of the cell where the fusion protein is most highly condensed, only the voxels in the top 5% of the fluorescence intensity distribution are shown in green. The lowest 5% are shown in red, highlighting primarily the boundary of the cell. This movie should be compared with Movie 2.

• Movie 4 -

  Movie 4. Light-adapted Xenopus rod expressing the arrestin-EGFP fusion protein Arr-EGFP rod. This rod is the same as the one in Movie 3 but the images were taken ~30 minutes after all rhodopsin was bleached and Arr-EGFP was redistributed between the IS and OS. The image uses a linear gradient of intensity display. Notice the banding pattern in the OS and also the ‘void’ at the base of the OS (the ellipsoid region), except for one side of the cell, where the ‘vestibule’ of the OS is seen.

• Movie 5 -

  Movie 5. Light-adapted Xenopus rod expressing the arrestin-EGFP fusion protein Arr-EGFP rod. This is the same rod cell as shown in Movie 3 and Movie 4, and the data are identical to those of Movie 4. However, in contrast to Movie 4 only the voxels in the top 5% of the fluorescence intensity distribution are shown in green, those in the lowest 5% are shown in red and all others were ‘turned off’ with Volview.