Fig. 2.

Validation of the Bag6–TRC35 interface. (A) Yeast two-hybrid assay to validate the interface identified in the crystal structure. WT TRC35 conjugated to the DNA binding domain was expressed with WT or mutant Bag6(951–1126) conjugated to the transcription activating domain. Interaction was determined by ability to grow on adenine-negative medium. (B) WT or mutant Bag6•GFP was coexpressed in Bag6^−/− 293T cells with TRC35•FLAG and immunoprecipitated using anti-GFP Ab. Amount of TRC35 retrieved by Bag6 was assessed by blotting with anti-FLAG Ab. The position of the higher molecular weight TRC35⋅FLAG is indicated by an asterisk.