Figure 2

Interaction of TACC1 and TRs in vitro , in yeast and in mammalian cells. (A) GST-pulldown assays were performed using in vitro translated [³⁵S]-TRα1 (left) or [³⁵S]-TRα2 (right) incubated with GST-glutathione-Sepharose (lanes 2 and 4), or GST-hTACC1-J-glutathione-Sepharose (lanes 3 and 5). Lanes 1 and 6 are [³⁵S]-TRα1 or [³⁵S]-TRα2 inputs (12.5%). (B, C) Flag-TACC1-J expression plasmid was cotransfected into COS-7 cells with TRα1, TRα2, TRβ1 (B) or TRα1 deleted from Helix12 expression plasmids (C), in the absence or presence of thyroid hormone [T3 (10^-7M)]. Forty-eight hours after transfection, whole cell lysates were prepared and immunoprecipitated with anti TR antibody. Immunoprecipitates were resolved by SDS-PAGE and blotted with anti-flag and anti-TR antibodies. Inputs correspond to 1% of the proteins used for the coimmunoprecipitation assay.