Talin and vinculin interact with the IDR of tensin3. (A) Western blot of tensin3 constructs N-terminally tagged with GFP expressed in NIH3T3 cells. (B) Co-expression of mCh-tensin3-IDR-cBAK with either GFP-vinculin or GFP-talin1 in NIH3T3 cells reveals either protein is recruited to the tensin3 IDR at mitochondria. Scale bar indicates 5 µm. (C) Secondary structure prediction of the tensin3 IDR (aa490-1120). Red boxes indicate the H1 and H2 regions. (D) mCh-talinFL-cBAK was expressed in NIH3T3 cells together with a tensin3-IDR construct lacking either the H1 (GFP-tensin3-IDR∆H1) or H2 (GFP-tensin3-IDR∆H2) motif. Note the absence of co-localization between GFP-tensin3-IDR∆H1 and mCh-talinFL-cBAK, indicating that the H1 motif is responsible for this interaction. (E) A tensin3 construct lacking the H1 motif (GFP-tensin3-FL∆H1) co-expressed in NIH3T3 cells with either mCh-vinFL-cBAK or mCh-talinFL-cBAK is not recruited to mitochondria under either condition. Scale bars in B and C indicate 10 µm. Source data are available for this figure: SourceData FS3.