Supplemental Material

This article contains the following supporting material:

• Fig2video1.mov - Control cells are motile. T47D control cells were plated on collagen (3μg/ml) for one hour and then a time-lapse sequence taken, with one image collected each minute for a duration of 90 minutes. All videos shown have a 30 frames/second display rate.
  
  
• Fig2video2.mov - Expression of activated R-Ras (38V) inhibits random migration. T47D cells stably expressing constitutively activated R-Ras (38V) were plated on collagen (3μg/ml) for one hour. A time-lapse sequence was collected, with one image taken per minute for 90 minutes.
  
  
• Fig2video3.mov - Cells expressing dominant negative R-Ras (41A) show enhanced migration. T47D cells stably expressing dominant negative R-Ras (41) were plated on collagen (3μg/ml) for one hour and then a time-lapse sequence taken, with one image collected each minute for 90 minutes.
  
  
• Fig3Video4.mov - Transfection of control siRNA oligos does not effect cell migration. T47D cells were transiently transfected with a pool of four non-specific control oligos. Two days later, the cells were plated on collagen (3μg/ml) for one hour and then a time-lapse sequence collected (one image collected each minute for a total of 90 minutes).
  
  
• Fig3Video5.mov - Knockdown of R-Ras blocks cell migration. T47D cells were transiently transfected with Alexa Fluor labeled-siRNA oligos specific for R-Ras. Two days later, the cells were plated on collagen (3μg/ml) for one hour and a time-lapse sequence collected. One image was collected per minute for 90 minutes. Every cell in the field is expressing the R-Ras siRNA, with the exception of the top cell (in the cluster of two cells) in the bottom right (see Figure 3).
  
  
• Fig8video6.mov - Inhibition of Rho does not rescue migration of R-Ras (38V) expressing cells. T47D cells stably expressing constitutively activated R-Ras (38V) were plated on collagen (3μg/ml) for one hour, treated with the Rho inhibitor, C3 exoenzyme (10μg/ml) for 10 minutes, and a time-lapse sequence collected. One image was collected per minute.
  
  
• Fig8video7.mov - Inhibition of ROCK rescues migration in cells expressing activated R-Ras (38V). T47D cells stably expressing constitutively activated R-Ras (38V) were plated on collagen (3μg/ml) for one hour, treated with the ROCK inhibitor, Y27632 (10μM) for 10 minutes, and a time-lapse sequence collected. One image was collected per minute.
  
  
• Fig10video8.mov - Expression of an effector mutant for R-Ras that uncouples binding to PI3-Kinase rescues migration. T47D cells stably expressing an effector mutant of constitutively activated R-Ras (38V/61S) were plated on collagen (3μg/ml) for one hour and then a time-lapse sequence was collected, with one image collected each minute for a total of 45 minutes.
  

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