Talin and vinculin do not interact at mitochondria. (A) The C-terminus of talin or vinculin was fused with the short mitochondrial targeting sequence from the outer mitochondrial protein BAK (cBAK). FAs are shown in green, actin stress fibers in orange, and mitochondria in blue. (B) When expressed in NIH3T3 cells, vinFL-cBAK and talinFL-cBAK both colocalize with the mitochondria-specific dye MitoTracker. Phalloidin staining showed no actin at the mitochondrial surface. (C) Staining with 9EG7 shows that activated integrins are absent from this system. Scale bars in A–C indicate 10 µm. (D) Coexpression in NIH3T3 cells of GFP-talinFL and (mCherry) mCh-vinFL-cBAK, or GFP-vinFL and mCh-talinFL-cBAK, reveals that full-length vinculin and talin do not interact with each other at mitochondria.