Biaxial Strain Induces FAK Activation

The biaxial strain was performed essentially as described elsewhere (Uzer et al., 2015 (link)). In brief, REF52 intact cells and cytoplasts were plated at a density of 30,000 cell/cm² per well in six-well BioFlex collagen-I coated plates (BF-3001C; Flexcell). After plating for either 3 or 18 h, cells were subjected to dynamic uniform biaxial cyclic strain at 5% magnitude at 10 cpm for 20 min using the Flexcell FX 5000 under conditions of 37°C and 5% CO₂. Control plates were handled the same but without strain application. Immediately after strain, whole-cell lysates were prepared and probed for phospho-FAK, total FAK, and GAPDH via Western blot analysis. Blots were analyzed using ImageJ. Data were derived from three independent experiments.