MBC Videos

This article contains the following supporting material:

• Fig3BD.mov (2.04 KB) - Dynamics of anti-zyxin mab 164D4 microinjected into REF-cell. While the Alexa488-coupled mAbs incorporate into stress fibers and focal adhesions, there is no recruitment of antibodies to the tips of the protruding lamellipodium (compare phase contrast).
  
  
• Fig3EF.mov (5.95 MB) - B16F1 cells expressing EGFP-zyxin compared with EGFP-VASP moving on laminin. Both constructs are targeted to focal adhesions. In addition, the motility of these cells induced on this substrate is accompanied by extensive recruitment of EGFP-VASP to the tips of protruding and ruffling lamellipodia. In contrast, EGFP-zyxin is not concentrated at these sites, although there is a slight increase in fluorescence intensity in the entire lamellipodium.
  
  
• Fig4.mov (5.80 MB) - Dynamics of EGFP-zyxin and rhodamine-labeled vinculin in B16-F1 cells on fibronectin. Vinculin and zyxin incorporate simultaneously into newly formed adhesions at the cell front (arrows). Conversely, zyxin dissociates from focal adhesions much earlier than vinculin (follow arrowheads). Focal adhesion disassembly is characterized in these cells by the detachment and sliding of vinculin-containing sites (arrowheads), which are devoid of zyxin.
  
  
• Fig5A.mov (5.97 MB) - In an experimental setup to initiate the sequence of events leading to focal adhesion disassembly, EGFP-zyxin-expressing CAR fibroblasts are injected with a mixture of constitutively active Rac and the Rho-kinase inhibitor Y27632. Under these conditions, zyxin is rapidly released from focal adhesions. The time-point of injection is indicated and can be followed in phase contrast.
  
  
• Fig5BC.mov (5.95 MB) - EGFP-Zyxin-expressing CAR cells already injected with fluorescent vinculin are injected again with the mixture of L61Rac/Y27632. Whereas zyxin dislocates from focal adhesions very rapidly, the distribution of vinculin remains almost unaffected. This experiment confirms the differential dynamics of vinculin and zyxin during adhesion disassembly observed in untreated cells (as shown in the supplemental video for Figure 4).
  
  
• Fig5FG.mov (6.08 MB) - EGFP-VASP-expressing CAR fibroblasts microinjected with rhodamine-labeled vinculin are injected again with the Rac/Y27632 mixture. In contrast with zyxin, this treatment causes a partial loss of VASP from focal adhesions, whereas the distribution of vinculin is again almost unchanged. Note that EGFP-VASP recruitment to the tips of lamellipodia is strongly enhanced by this treatment.
  

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