FIGURE 4.

JKAP dephosphorylates FAK and decreases paxillin-rich focal adhesions. A, H1299-JKAP_TR cells and H1299-JKAP-CS_TR cells were transfected with Src and then incubated with 2 μg/ml tetracycline (Tet) for 6 and 12 h. The cell extracts were subjected to Western blotting using specific antibodies as indicated. Results shown are one representative result from three independent experiments. The relative band intensities on Western blot assay shown under blot were determined using a computing densitometer equipped with the Gel-Pro analyzer program, which were normalized by arbitrarily setting the densitometry of control to 1. B, bar graphs depicted the results from A (open bars, H1299-JKAP_TR; shaded bars, H1299-JKAP-CS_TR cells). Quantitative data were means ± S.E. of three independent experiments. *, p < 0.05; **, p < 0.01, compared with Src-only control. C, H1299-JKAP_TR cells were cultured with or without 2 μg/ml tetracycline for 12 h and subjected to double immunostaining with an anti-paxillin antibody (red) as well as an anti-Myc antibody (green). Cell nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI) (blue). Arrows indicate those cells with higher JKAP expression and less paxillin-rich focal adhesions.