MBC Videos

This article contains the following supporting material:

• video1.avi (4.06 MB) - Cos7 cells 36hr after transfection with GFP-tagged WIPI49. The cell shown has a moderate level of overexpression. This video shows the migratory behaviour of the peripheral membranes positive for GFP-WIPI49. These membranes can be seen to undergo both short- and long-range bidirectional movements. Playback speed is 3 times actual.
  
  
• video2.avi (9.91 MB) - Cos7 cells 36hr after transfection with GFP-tagged WIPI49. The cellular region shown is typical of the perinuclear fluorescence observed in approximately 30% of the GFP-WIPI49 transfectants. It is notable that in the time frame observed that there is very little traffic between the peripheral endosomal pool of GFP-WIPI49 and the Golgi localised pool. Playback speed is 3 times actual.
  
  
• video3.avi (2.14 MB) - Cos7 cells were transfected with plasmids encoding pEGFPTubulin (blue) and pDsRED2-WIPI49 (red). 36 hours post-transfection cells were filmed and data recorded over 587 seconds. A number of DsRED2-WIPI49 positive structures can be seen to migrate along microtubules. This is particularly clear in the boxed area indicated. Playback speed is 22 times actual.
  
  
• video4.avi (5.22 MB) - Supplement to Figure 5. Cos7 cells 36hr after transfection with DsRED2-tagged WIPI49 (red) and GFP-Rab5 (blue). In the cell shown there are a number of regions with a partial colocalisation and juxtaposition between the two proteins. These structures can be seen to move together in a coordinate fashion and in some cases segregate from each other. The boxed region in the upper right hand corner of the cell (shown in Figure 5) refers to a specific event wherein Rab5 structures can be seen to migrate to, associate with, and then depart from WIPI49- positive membranes. This is more clearly shown in the gallery depicted in Figure 5B. Playback speed is 8 times actual.
  
  
• video5.avi - Cos7 cells 36hr after transfection with DsRED2-WIPI49 (red) and GFP-Rab7 (blue). In the cellular region shown it is apparent that whilst some of theRab7 and WIPI49 structures appear to undergo coordinate movement, there is minimal colocalisation between the proteins. Playback speed is 21 times actual.
  
  
• video6.avi (1.82 MB) - Cos7 cells 36hr after transfection with DsRED2-WIPI49 (red) and GFP-Rab11 (blue). There is no apparent relationship between the ectopic WIPI49 and Rab11, neither in terms of colocalisation nor coordinate movement of membranous structures. Playback speed is 21 times actual.
  
  
• video7.avi (2.76 MB) - Supplement to Figure 6. Cos7 cells 36hr after transfection with DsRED2-WIPI49 (red) and YFP-Rab9 (blue). In the region of the transfected cell shown there is a substantial level of colocalisation and juxtaposition between the two ectopic proteins. These structures can be seen to move in a coordinate fashion together. Importantly one can observe WIPI49 vesicles migrating to and associating with Rab9 elements. This point is highlighted for one event in particular by the boxed region of Figure 6A and in the gallery of Figure 6B. The whole film represents approximately 300 seconds in real time and playback speed is 12 times actual.
  
  
• video8.avi (4.97 MB) - Supplement to Figure 9. Cos7 cells 36hr after transfection with GFP-WIPI49 were treated with LY294002. Cells were filmed for twenty minutes after addition of drug. The membrane-associated fluorescence diminishes over time. Playback speed is 97 times actual.
  
  

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