Tensin3 binds multiple talin rod domains. (A–C) Western blotting of the mitochondrial pulldown experiments in HEK293T cells with the same constructs used in Fig. 1, G–I, showing that R3, R4, and R11 interact with mCh-IDR. (D) Representative images of NIH3T3 cells co-expressing mCh-TNS3-cBAK with GFP-vector, GFP-R1R2, GFP-R1R3, GFP-R4R6, GFP-R5R6, GFP-R7R8, GFP-R9R10, GFP-R11DD, or GFP-R12DD, respectively. Black boxes indicate colocalization, and gray boxes indicate no association. Note that TNS3-cBAK recruits R1R3, R4R6, R7R8, and R11DD to the mitochondria, but not GFP-vector, R1R2, R5R6, R9R10, or R12DD. (E) Summary table of the MTS experiments in D. (F–I) Representative images of NIH3T3 cells co-expressing GFP-R1R3-cBAK (F), GFP-R4R6-cBAK (G), GFP-R7R8-cBAK (H), or GFP-R11DD-cBAK (I) with mCh-IDR-WT or mCh-IDR-L702E, respectively. Note that L702E abolishes mCh-IDR interactions with all talin1 truncation constructs. (J) Images of cells co-expressing mCh-TNS3-cBAK (WT or L702E) with GFP-TLN1-E1770A, respectively. L702E abolishes TNS3 colocalization with TLN1-E1770A at the mitochondria. All experiments are performed three times. Scale bars, 5 μm. Source data are available for this figure: SourceData FS1.
Figure S1.

Tensin3 binds multiple talin rod domains. (A–C) Western blotting of the mitochondrial pulldown experiments in HEK293T cells with the same constructs used in Fig. 1, G–I, showing that R3, R4, and R11 interact with mCh-IDR. (D) Representative images of NIH3T3 cells co-expressing mCh-TNS3-cBAK with GFP-vector, GFP-R1R2, GFP-R1R3, GFP-R4R6, GFP-R5R6, GFP-R7R8, GFP-R9R10, GFP-R11DD, or GFP-R12DD, respectively. Black boxes indicate colocalization, and gray boxes indicate no association. Note that TNS3-cBAK recruits R1R3, R4R6, R7R8, and R11DD to the mitochondria, but not GFP-vector, R1R2, R5R6, R9R10, or R12DD. (E) Summary table of the MTS experiments in D. (F–I) Representative images of NIH3T3 cells co-expressing GFP-R1R3-cBAK (F), GFP-R4R6-cBAK (G), GFP-R7R8-cBAK (H), or GFP-R11DD-cBAK (I) with mCh-IDR-WT or mCh-IDR-L702E, respectively. Note that L702E abolishes mCh-IDR interactions with all talin1 truncation constructs. (J) Images of cells co-expressing mCh-TNS3-cBAK (WT or L702E) with GFP-TLN1-E1770A, respectively. L702E abolishes TNS3 colocalization with TLN1-E1770A at the mitochondria. All experiments are performed three times. Scale bars, 5 μm. Source data are available for this figure: SourceData FS1.

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