JCS03167 Supplementary Material

Files in this Data Supplement:

• Movie 1 -

  Movie 1. EGF stimulation rapidly induces highly dynamic SNX5-positive tubule formation on newly formed macropinosomes. Images were taken every 10 seconds over a time period of 27 minutes. Tif stacks were converted to QuickTime format using ImageJ (NIH). The movie is played at 20 frames/second. Bar, 10 μm.

• Movie 2 -

  Movie 2. SNX5-positive compartment is accessible to fluid-phase markers. Images were taken every 10 seconds over a time period of 5 minutes. RGB stacks were converted to QuickTime format using ImageJ (NIH). The movie is played at 20 frames/second. Bar, 10 μm.

• Movie 3 -

  Movie 3. SNX5-positive tubule extension and departure plays a role in the maturation of the macropinosome. Images were taken every 10 seconds over a time period of 15 minutes. Tif stacks were converted to QuickTime format using ImageJ (NIH). The movie is played at 20 frames/second. Bar, 10 μm.

• Movie 4 -

  Movie 4. Extension of SNX5-positive tubules is dependent on microtubules. Images were taken every 10 seconds over a time period of 15 minutes. Tif stacks were converted to QuickTime format using ImageJ (NIH). The movie is played at 20 frames/second. Bar, 10 μm.

• Movie 5 -

  Movie 5. Macropinosome-derived SNX5-positive tubules traffic to smaller SNX5-positive early endosomes.Images were taken every 1 second over a time period of 15 minutes. Tif stacks were pseudocoloured to highlight variations in fluorescent intensity (black/blue, low intensity; white/yellow, high intensity) and converted to QuickTime format using ImageJ (NIH). The movie is played at 20 frames/second.

• Movie 6 -

  Movie 6. Macropinosome-derived SNX5-positive tubules traffic to smaller SNX5-positive early endosomes. Images from Movie 5 were cropped and stacked on the Z-axis using software developed for the task. The frames were arranged with the first frame in the foreground with each successive frame behind the first. Presented is a rotation of the temporal construct. Labels indicate frame number.

• Movie 7 -

  Movie 7. SNX5-positive tubule departure is concurrent with Rab5 to Rab7 conversion. Images were taken every 2 seconds over a time period of approximately 5 minutes. Tif stacks were converted to QuickTime format using ImageJ (NIH). The movie is played at 20 frames/second. Bar, 10 μm.

• Movie 8 -

  Movie 8. SNX5-positive tubule departure is concurrent with Rab5 to Rab7 conversion. Images were taken every 2 seconds over a time period of approximately 5 minutes. Tif stacks were converted to QuickTime format using ImageJ (NIH). The movie is played at 20 frames/second. Bar, 10 μm.