JCS01398 Supplementary Material

Files in this Data Supplement:

• Movie 1
• Movie 2 -

  Movies 1 and 2. Recruitment of PH-Akt-GFP to the cell’s leading edge in transiently transfected HEK 293 cells stably expressing wild-type CXCR2 or CXCR2 mutated in the AP-2 binding motif, respectively, in response to a gradient delivery of MIP-2. The arrows at the beginning and end of the movies indicate the direction of gradient. Images were taken every minute and are displayed at three frames/second.

• Movie 3
• Movie 4 -

  Movies 3 and 4. Localized activation of Rac1, and FRET analysis in transiently transfected HEK 293 cells with stable expression of wild-type CXCR2 or CXCR2 mutated in the AP-2 binding motif, respectively, in response to a gradient delivery of MIP-2.

• Movie 5
• Movie 6 -

  Movies 5 and 6. Localized activation of Cdc42 with FRET analysis in transiently transfected HEK 293 cells stably expressing wild-type CXCR2 or CXCR2 mutated in the AP-2 binding motif, respectively, in response to a gradient delivery of MIP-2. FRET images were made as ratio images from two different fluorescent images (YFP/CFP) and plotted with pseudocolor. Red, highest activity; blue, lowest activity. Each pair of images was taken every 30 seconds and is displayed at five frames/seconds for movies 3-6.

• Movie 7
• Movie 8 -

  Movies 7 and 8. Intensity increase of Cdc42 FRET signal activated with a gradient of MIP-2 was inhibited by the expression of either dominant negative (Movie 7) or constitutive active Cdc42 (Movie 8). MIP-2 gradient was delivered in the direction shown with arrow in the first frame of the movies.