Rabbit anti phospho fak

The following antibodies were used: Rabbit anti-phospho-FAK (Tyr925, Cell Signaling Technologies, Danvers, MA, USA), purified Mouse anti-N-cadherin (BD Biosciences, New Jersey, USA), Mouse anti-E-cadherin (Invitrogen, Life Technologies, Monza, Italy), Mouse anti-VEGFR2 (R&D, Minneapolis, MN), β-Actin HRP-conjugated (Sigma, Milano, Italy), stabilized Goat anti-Mouse HRP-conjugated (Thermo Fisher Scientific, Waltham, MA), stabilized peroxidase conjugated Goat anti-Rabbit [(H + L), Thermo Fisher Scientific]. Reagents used are: VEGF-A165 (Peprotech, Rocky Hill, NJ, USA), trans-Ned-19 (Tocris Bioscences, Bristol, United Kingdom), Bafilomycin A1 (Sigma), Thapsigargin (Sigma). Ned-19 was purchased from Tocris Bioscience and was at least 98% pure. Methanol and other organic solvents were chromatography grade from Sigma, water was purified using a milliQ system from Millipore (Billerica, MA, USA) and LC-MS grade formic acid was obtained from Fluka (Sigma). Centrifugal ultrafiltration devices were Amicon Ultra-0.5 (Ultracel-3 K, regenerated cellulose 3,000 MW) from Millipore Co. Ltd.