FIGURE 1.

Paxillin negatively regulates the anchorage-independent growth of HeLa cells. A, images of HeLa cells that had been either mock-transfected or transfected with GFP-Pxn WT, as indicated, and then stained with GFP and Cat-1 antibodies. Arrows indicate some of the focal complexes where both GFP-tagged Pxn WT and Cat-1 were detected. B and C, two sets of HeLa cells were transfected with either Cont. siRNA or Pxn siRNA 1 or Pxn siRNA 2. One set (B) was lysed and then immunoblotted with Pxn and Cat-1 antibodies. Soft agar assays were performed (C) on the other set of cells. The number of colonies that formed for each condition was determined. D and E, two sets of NIH3T3 cells stably expressing an activated form of Cdc42, Cdc42 F28L, were transfected with either Cont. siRNA or Pxn siRNA 1 as indicated. One set (D) was lysed and then immunoblotted with Pxn and Cat-1 antibodies. Soft agar assays were performed (E) on the other set of cells. The number of colonies that formed for each condition was determined. All of the experiments shown were performed three separate times, each yielding similar results. The data shown in C and E represent mean ± S.D. Student's t tests were performed. *, p < 0.05.