JCS028738 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. The integrin-mediated conformational change in the FERM domain of adherent cells is not influenced by K38A mutation. (A) Acceptor photobleaching FRET efficiency (E%) images of FAK^−/− fibroblasts expressing the FERM sensor (upper panels) or K38A mutant of the FERM sensor (lower panels), replated on PLO for 30 minutes or adherent on fibronectin (FN). YFP was bleached in the boxed area, FRET efficiencies are shown in pseudo-color. Bars, 10 µm. (B,C) Cumulative (left) and integrated cumulative (right) FRET distributions of the cytosol and FAs of (B) PLO-plated and (C) fibronectin-adherent cells expressing the K38A mutant as compared to the native FERM sensor. KS testing revealed that the K38A mutation significantly increased the sensor response in PLO-plated cells (P<0.001). Although the K38A mutation is able to destabilize the FERM conformation, no difference is observed for integrin-mediated FERM activation of FAK in fibronectin-adherent cells.

• Supplemental Figure S2 -

  Fig. S2. Dynamic FA behaviors in randomly migrating FAK^−/− fibroblasts. Examples of FA behaviors in randomly migrating FAK^−/− fibroblasts expressing the FERM sensor and imaged at 5-minute intervals (A) Growing (arrow 1), shrinking (arrows 2), and (B) stable (arrow 3) FAs are shown. The YFP fluorescence intensity distributions (YFP), red/green image overlays of different times, and corresponding FRET ratios (YFP/CFP) are shown in pseudo-color as indicated. Bars, 10 µm.