JCS01531 Supplementary Material

Files in this Data Supplement:

• Movie 1 -

  Movie 1. Time-lapse sequence of a fibroblast expressing GFP-tau. Images were acquired at 3-second intervals over a 4.5-minute period and are played back at 10 frames/second.

• Movie 2 -

  Movie 2. Time-lapse sequence of GFP-tau-decorated MTs at the lamella region of a fibroblast. Images were acquired at 3-second intervals over a 1.5-minute period and are played back at 10 frames/second.

• Movie 3 -

  Movie 3. Time-lapse sequence of a fibroblast expressing GFP-tau after addition of 1 mM taxol. Images were acquired at 3-second intervals over a 3-minute period and are played back at 10 frames/second. Notice rapid decrease in the amount of GFP-tau associated with MTs. Photobleaching is negligible during the time course of the sequence.

• Movie 4 -

  Movie 4. Time-lapse sequence of a human 356 fibroblast expressing GFP-tau after addition of 1 mM taxol. Images were acquired at 3-second intervals over a 5-minute period and are played back at 10 frames/second. Notice the rapid decrease in the amount of GFP-tau associated with individual MTs at the cell periphery. Photobleaching is negligible during the time course of the sequence.

• Movie 5 -

  Movie 5. Time-lapse sequence of a fibroblast expressing GFP-tau after extraction in a buffer supplemented with 10 mM taxol. Images were acquired at 1-second intervals over a 2.5-minute period and are played back at 10 frames/second. The shift of focus at the beginning of the sequence is because of the addition of the buffer. Extraction of the plasma membrane led to a rapid decline in the cytoplasmic fluorescence because of the loss of free GFP-tau. Within seconds of buffer application, most of the GFP-tau dissociates from the MTs.

• Supplemental Figure 1 -

  Fig. S1. Phosphatase inhibitor calyculin A induces dissociation of tau from individual MTs in Xenopus neurons. A series of fluorescence micrographs of the flat axonal segment of GFP-tau-expressing neuron in the vicinity of the branch point. Time after calyculin A application (200 nM) is in seconds at the upper-left corner of each image. Individual tau-decorated MTs (arrows) are clearly visible before and 60 seconds after calyculin A application. MTs are more difficult to detect at later time points. Progressive increase in the background fluorescence after calyculin A application reflects higher concentration of free cytoplasmic GFP-tau because of its dissociation from MTs. Bar, 10 mm.