Figure 2.
Active vinculin can bind talin independently of force. (A) Coexpression of active mCh-vinT12-cBAK with GFP-talinFL in NIH3T3 cells shows that the two constructs colocalize at mitochondria. (B) This interaction occurs in the presence of Y-27632 (50 µM), blebbistatin (50 µM), or cytochalasin D (Cyto D; 2.5 µg ml−1). (C) mCh-vinT12-cBAK also recruited a talin construct that has mutations in both actin binding sites in the talin rod (ABS2 and ABS3; GFP-talinABS2+ABS3mut) in NIH3T3 cells. Scale bars in A–C indicate 10 µm. (D) FLAP experiments in NIH3T3 cells coexpressing mCh-vinT12-cBAK and photoactivatable (PA) GFP-talinFL show that there is minimal loss of fluorescence over time after activation, indicating a very strong interaction between the two proteins. Error bars represent SEM; n = 11 mitochondria from 5 cells. Results are representative of three independent experiments. Scale bar indicates 5 µm.

Active vinculin can bind talin independently of force. (A) Coexpression of active mCh-vinT12-cBAK with GFP-talinFL in NIH3T3 cells shows that the two constructs colocalize at mitochondria. (B) This interaction occurs in the presence of Y-27632 (50 µM), blebbistatin (50 µM), or cytochalasin D (Cyto D; 2.5 µg ml−1). (C) mCh-vinT12-cBAK also recruited a talin construct that has mutations in both actin binding sites in the talin rod (ABS2 and ABS3; GFP-talinABS2+ABS3mut) in NIH3T3 cells. Scale bars in A–C indicate 10 µm. (D) FLAP experiments in NIH3T3 cells coexpressing mCh-vinT12-cBAK and photoactivatable (PA) GFP-talinFL show that there is minimal loss of fluorescence over time after activation, indicating a very strong interaction between the two proteins. Error bars represent SEM; n = 11 mitochondria from 5 cells. Results are representative of three independent experiments. Scale bar indicates 5 µm.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal