Immunodetection of Focal Adhesion Proteins

The following antibodies were used: anti-YAP/TAZ (sc-101199, Santa Cruz Biotechnology), anti-tensin-1 (SAB4200283, Sigma-Aldrich), anti-fibronectin (F3648, Sigma-Aldrich), anti-vinculin (clone hVIN-1, V9131), anti-talin (clone 8d4, T3287, Sigma-Aldrich), anti-paxillin (612405, BD Biosciences), anti-phosphotyrosine (Y118)-paxillin (2541, Cell Signaling Technology), anti-phosphotyrosine (Y397)-FAK (clone D20B1, 8556, Cell Signaling Technology) and anti-tensin-3 (ABT29, Millipore). Anti-human active α5-integrin (SNAKA51) was a gift from Martin Humphries, University of Manchester, UK (Clark et al., 2005 (link)). Phalloidin Atto 647N (65906) was obtained from Sigma-Aldrich. Alexa Fluor^®-conjugated secondary antibodies (Alexa Fluor 488, 555, 568, 647-conjugated anti-mouse-, rabbit- and rat-IgG antibodies, Thermo Fisher Scientific) were used in immunofluorescence. Bovine plasma fibronectin (FN; 341631) was purchased from Merck, Millipore and collagen type I (from calf skin) from Sigma-Aldrich (C8919).