Figure 3.

Hr interacts directly with TR. (A) Far Western assay for interaction of Hr with TR. Extracts from bacteria expressing trpE-Srg1 (negative control), trpE-hr, GST only, and GST-hr were separated by SDS/PAGE and transferred to nitrocellulose. Total protein was detected by amido black staining (Left); Hr fusion proteins were detected by Western blot analysis with anti-hr antisera (Center); interaction of Hr with TR was detected by far Western blot analysis with ³⁵S-labeled TRβ1 (Right). Detection of multiple lower molecular mass bands by [³⁵S]TR is from binding to fusion protein degradation products. Sizes of molecular mass standards are in kilodaltons. (B) Effect of hormone on TR/Hr interaction. Far Western assay was as described above with ³⁵S-labeled TRβ1 in the absence (Left) and presence (Right) of thyroid hormone.