FIGURE 7.
Mutations of the FAT H1-H2 hinge alter interaction of FAK with paxillin but not talin. A, COS7 cells were transfected with VSV-tagged WT FAK, R-FAK, or T-FAK, without or with B-Fyn cotransfection, as indicated. Cells were lysed 24 h later and FAK immunoprecipitated with anti-VSV antibodies (IP VSV). Phosphorylated Tyr925 was determined in total cell lysates (pY925, upper panel). Expression of B-Fyn was detected by immunoblotting. Immune precipitates were probed with FAK, paxillin, and talin antibodies as indicated (lower panels). Mass markers positions are indicated in kDa. B, quantification of the co-immunoprecipitation of paxillin with FAK as in A expressed as a ratio of paxillin/FAK, a.u., arbitrary units. Statistical analysis, one-way ANOVA: F(5,18) = 18.09, p < 0.0001, Tukey's post hoc test; *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001. C, quantification of the co-immunoprecipitation of talin with FAK as in A expressed as a ratio of talin/FAK, a.u., arbitrary units. One-way ANOVA: F(5,18) = 0.35, p = 0.87. B and C, data are mean ± S.E.