JCS03321 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. (A) STICS velocity map generated from a computer simulated image time series with simulated adhesion areas having an immobile particle population (80% of the particles) and a flowing population (20% of the particles |v_y|=−0.1 μm/second). Off adhesion particle populations were a factor of six less bright and were immobile (80% of the particles) or diffusing (20% of the particles D=1.0 μm² /second). Spatial scale bar is 5 μm, velocity scale arrow is 1 μm/minute. (B) STICS velocity maps calculated from regions encompassing a long adhesion (20×5 pixels, white rectangle), adhesion plus some surrounding area (16×16 pixels,red square), and an off adhesion area (16 x 16 pixels, white square) for a MEF cell transiently expressing EGFP-talin. Scale bar is 5μm and velocity scale bar is 0.2 μm/second.

• Supplemental Figure 2 -

  Fig. S2. Velocity maps for talin-EGFP and mRFP-actin in a retracting region of a CHO cell plated on 2 μg/ml fibronectin. The color-coding for the velocity vectors is universal with blue being slow velocities and red being faster velocities. The scale of the velocity vectors is different for each plot and is referenced on each image with a 0.5 μm/minute velocity scale arrow. Spatial scale bars are 5 μm. Pixel size is 0.215 μm.

• Movie 1A
• Movie 1B -

  Movie 1. Movies of α-actinin (a) and actin (b) in MEF cells plated on 1 μg/ml fibronectin from Fig. 3A. Time stamp is in seconds. Note that the analysis software rotates and inverts the images so they are not oriented the same way in the figures and the movies.

• Movie 2A
• Movie 2B -

  Movie 2. Movies of α-actinin in a CHO cell plated on 2 μg/ml fibronectin from Fig. 4A both before (a) and after (b) treatment with 200 nM cytoD. Time stamp is in seconds. Note that the analysis software rotates and inverts the images so they are not oriented the same way in the figures and the movies.

• Movie 3A
• Movie 3B
• Movie 3C
• Movie 3D -

  Movie 3: Movies of paxillin (a) and actin (b) or talin (c) and actin (d) in CHO cells plated on 2 μg/ml fibronectin from Fig. 5A. Time stamp is in seconds. Note that the analysis software rotates and inverts the images so they are not oriented the same way in the figures and the movies.

• Movie 4A
• Movie 4B -

  Movie 4. Movies of paxillin (a) and actin (b) in a 3T3 cell plated on 1 μg/ml fibronectin from Fig. 6A. Note the more highly organized actin filaments and larger focal adhesions relative to supplementary Movies 3a and b in CHO cells. Time stamp is in seconds. Note that the analysis software rotates and inverts the images so they are not oriented the same way in the figures and the movies.

• Movie 5A
• Movie 5B -

  Movie 5. Movies of talin (a) and actin (b) in a CHO cell plated on 5 μg/ml fibronectin from Fig. 7A. Time stamp is in seconds. Note that the analysis software rotates and inverts the images so they are not oriented the same way in the figures and the movies.

• Movie 6A
• Movie 6B -

  Movie 6. Movies of talin (a) and actin (b) in a retracting region of a CHO cell plated on 2 μg/ml fibronectin from supplementary Fig. S1. Time stamp is in seconds. Note that the analysis software rotates and inverts the images so they are not oriented the same way in the figures and the movies.