Supplemental Material

This article contains the following supporting material:

• Supplemental Figure 1 - 2-3-2-RFP interacts selectively with p190RhoGAP. (Top) HEK293 cells were transfected with the phosphotyrosine-containing proteins p190RhoGAP, p120RasGAP, Paxillin (Pax), Src, or Cortactin (Cort) and either RFP or 2-3-2-RFP and Arg. The phosphotyrosine-containing proteins were immunoprecipitated and probed for RFP to detect coimmunoprecipitating RFP or 2-3-2-RFP. The same blot was probed for each protein to ensure proper immunoprecipitation (IP panels). Total protein lysates (25 μg) were probed for the transfected proteins (Total Protein panels).

• Supplemental Figure 2 - p190 and Arg do not colocalize at the cell periphery during cell spreading. Arg^–/– + Arg-YFP cells (A, D, G) were plated on FN-coated coverslips for 10, 20, or 30 minutes, then fixed, and stained for p190RhoGAP (B, E, H). Merged images (C, F, I) show Arg in green and p190 in red. Scale bar in A, 10 μm.