JCS02693 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig.S1. PTP-PEST^–/– cells were transfected with the indicated ratios of PTP-PEST:pCDNA3.1 empty vector (total of 1mg).  Cells were spread on 10 µg/ml fibronectin for the times indicated. Quantitation of inhibition of spreading was measured as the percentage of round GFP-positive cells. 

  
• Movie 2a
• Movie 2b
• Movie 2c -

  Fig.S2. Paxillin^–/– cells were transfected with GFP, PTP-PEST plus wild-type paxillin, or PTP-PEST plus paxillin C523S mutant.  Cells were replated on 10 mg/ml fibronectin-coated dishes and spread for three hours at 37°C for 3 hours. GFP-positive cells were identified and time-lapse images were acquired every 150 seconds for 120 minutes. 

• Supplemental Figure 3 -

  Fig.S3. Paxillin^–/– cells transfected with GFP and paxillin constructs (in the absence of PTP-PEST overexpression) as indicated, were plated on 10 mg/ml fibronectin-coated dishes for 180 minutes followed by acquisition of time lapse images every 10 minutes.  Changes in cell protrusion area of at least 10 transfected cells were quantified at 10-minute intervals for 1 hour as described in Materials and Methods.  *P< 0.05.  Paxillin alone significantly inhibited cell protrusion.  The expression of paxillin DLD4 and Y31/118F, modestly suppressed cell protrusion, whereas 31/118FDLD4 had no effect

• Supplemental Figure 4 -

  Fig.S4. PTP-PEST^–/– cells were transfected with various concentrations of PTP-PEST cDNA as indicated. Modified Boyden chamber assays were performed to evaluate the optimal rescue of migration.