Pf 573228

The following monoclonal antibodies were used: mouse AC-15 to actin (Sigma, München, Germany), mouse B-1 to Akt1 (Santa Cruz, Heidelberg, Germany), rabbit D9E to phospho-Akt (S473) (Cell signaling technology, Danvers, USA), mouse 36/E-cadherin (BD Bioscience, Heidelberg, Germany), mouse 4.47 to phospho-FAK Y397 (Millipore, Darmstadt, Germany) and rat MB1.2 to β1 integrin (Merck, Darmstadt, Germany). The polyclonal antibodies were: rabbit ERK2 (C-14), rabbit GAPDH and rabbit integrin β4 (H-101) (Santa Cruz, Heidelberg, Germany), rabbit phospho-p44/42 MAPK (ERK1/2; T202/Y204) (Cell signaling technology, Danvers, USA), rabbit phospho-FAK (Y397) (Invitrogen, Karlsruhe, Germany), rabbit Keratin 5 (HISS Diagnostic, Freiburg, Germany) and the rabbit Endo-2 antibody to ColXVII [15] (link).
A polyclonal rabbit antibody specific for the phosphorylated residue S1356 on the β4 integrin subunit was kindly provided by A. Sonnenberg (The Netherlands Cancer Institute) [16] .
The inhibitor PI3K (LY294002) was obtained from Merck Biosciences (Darmstadt, Germany). Selective focal adhesion kinase (FAK) inhibitors PF 573228 and FAK Inhibitor 14 were obtained from R&D Systems (Wiesbaden-Nordenstadt, Germany). Recombinant EGF was obtained from ImmunoTools (Friesoythe, Germany). All inhibitors are used in indicated concentrations.

The following inhibitors or activators were used for signaling studies: 1–5 mM EDTA, 10 µM PP2, 1 µM colchicine (Sigma), 6 mM acrylamide (National Diagnostics, Atlanta, GA, USA), 1 µM AG82 FAK/Src inhibitor, 3 µM manumycin (Calbiochem, San Diego, CA, USA), 500nM PF 573228 (R&D Systems, MN, USA), and 5 µM LPA (Enzo Life Sciences).