TORCAR activity in FAs and adjacent regions upon FCS stimulation. Representative confocal time-lapse video of GFP-paxillin intensity-scaled ratio image of TORCAR FRET-based biosensor in serum-starved HeLa cells (left). The movie highlights the normalized ratio change as well as the intensity of the biosensor in different parts of the cells during a 13-min measurement (80 frames in total, 10-s interval). The traces of the normalized ratio change (right) are shown with the same color coding as the ROIs in the image. Cold-colored ROIs correspond to FAs and warm-colored ROIs to control areas (adjacent to FAs). The cells were treated after 3 min with 10% FCS until the end of the measurement. Upon stimulation, the normalized ratio within the ROIs showing FAs decreases strongly, which is visualized by red color. The quantification of such measurements is shown in Fig. 3 and Fig. S3.
