Volume 8 issue 7 July 2007
Microtubules Tethered at Epithelial Cell Junctions by Dynein Facilitate Efficient Junction Assembly
LA Ligon and ELF Holzbaur

Supplemental Figure 1.  PtK2 epithelial cells were transfected with a plasmid encoding GFP-tubulin, using conditions that allowed a low level of expression in about 20% of the cells.  Cells were placed in a heated recording chamber and fluorescent images were taken every ~10 seconds for 5-6 minutes.  Shown here is one cell expressing GFP tubulin making contact with a neighboring cell that does not express labeled tubulin.

Video 1 (mov)

Supplementa Figure 2. PtK2 epithelial cells were transfected with a plasmid encoding GFP-tubulin, then treated with 4mM EGTA for 20 minutes to disassemble cell-cell junctions. Cells were then allowed to recover in calcium-containing media.  During recovery, cells were placed in a heated recording chamber and images were taken every ~10 seconds for 10-12 minutes.  Shown here is a time-lapse video of the microtubule behavior with several microtubules pseudocolored to highlight their activities.  

Video 2 (mov)