Talin contains tensin-binding sites in the R3, R4, R7R8, and R11 domain. (A) Schematic representation of talin with the aa numbers of each domain indicated. (B) Schematic representation of tensin3. Talin-binding site (TBS), aa 692–718. (C) Mitochondrial targeting talin1 full-length and deletion constructs. The indicated talin sequences were inserted between EGFP (GFP) and cBAK, with the aa number of talin regions indicated below. (D) Constructs shown in C were expressed with mCh-IDR in NIH3T3 cells. Black boxes indicate colocalization, and gray boxes indicate no association. (E) Western blotting of mitochondrial pulldown experiments. Constructs as used in D were expressed in HEK293T cells. Whole cell lysates (wcl) and purified mitochondria were immunoblotted. Note that the double band for mCh-IDR is due to known mCherry degradation. (F) Representation of additional talin1 constructs. (G–I) NIH3T3 cells expressing the constructs shown in C and F with mCh-IDR. (J) Summary table of the mitochondrial targeting assays. All results are collected from three independent experiments. Scale bars (D and G–I), 5 μm. aa, amino acid. Source data are available for this figure: SourceData F1.