Figure 6

Pak1 is associated with focal adhesions. BxPC-3 cells were stimulated with 10 nM of mature TGFβ₁ for ten minutes followed by preparation of the Triton-X100 nonsoluble fraction and precipitation with α_Vβ₆ integrin antibodies. The precipitate was then reprecipitated with anti-FAK antibodies (sc-1688), followed by re-precipitation with anti-phospho^S (ab9334, Abcam), anti-phospho^Thr (ab2286), and analyzed with a Pak1 antibody (71-9300, Zymed). In part, the cells were preincubated with 50 μM PP2 (Calbiochem), and 50 μM PD98059 (Calbiochem), respectively [10].