Fak inhibitor 14 fak 14

DAPI (Invitrogen D1306) was dissolved at 5mg/mL in dimethyl formamide and used at 1/5000. Other drugs were dissolved in DMSO (endotoxin-free dimethyl sulfoxide, Sigma D2650) at stock concentrations indicated below. Stock concentrations and sources of drugs were: 50mM LY294002 (Sigma L9908), 25mM wortmannin (EMD Chemicals 12–338), 30mM Y27632 (EMD Chemicals 688000), 10mM FAK inhibitor-14 (FAK-14) (Tocris Bioscience 3414), 100mM PF573228 (Tocris Bioscience 3239). SMIFH2 (Millipore 344092) solutions were freshly made with each experiment as we found that frozen stocks degraded over time. Primary antibody used for inside/outside staining was BacTrace anti-Listeria genus specific antibody (01-90-90, KPL, Inc.). Fluorescent streptavidins used for inside/outside staining of 2.0μm biotinylated polystyrene beads (Polysciences, Inc. 24172) were Alexa-Fluor-546-streptavidin (Invitrogen S11225) and Alexa-Fluor-488-streptavidin (Invitrogen S11223). For Western blotting, rabbit monoclonal anti-Arp2 antibody (Epitomics 5738–1) was used to detect Arp2.

The ROCK inhibitors Y-27632 (Rho Kinase inhibitor VI) and H-1152 were obtained from EMD Millipore (Billerica, MA, USA); Q-VD-OPh from MP Biomedicals (Eschwege, Germany); and GT from AppliChem (Darmstadt, Germany). The CNFy toxin (150 ng/ml) from Yersinia pseudotuberculosis and the C2I/C3 fusion toxin (Clostridium botulinum and Clostridium Limosum, respectively) combined with the C2II toxin from Clostridium botulinum (collectively termed C3 toxin) was produced and purified as described^{33 (link),34 (link)}. C3 toxin is a binary toxin consisting of C2II (200 ng/ml) and C2I/C3 (100 ng/ml) mixed in cell culture media. The FAK inhibitor 14 (FAK14) was purchased from Tocris (Bristol, UK), and Cilengitide, a cyclic pentapeptide RGD compound (cyclo-[RGDfN(Me)V])^{46 (link)}, the RGD peptide, DTT and iodocetamide were from Sigma (Taufkirchen, Germany).