Transcriptional regulation controlled by thyroid hormone receptor (TR) drives events such as development, differentiation and metabolism. TRs may act either as homodimers, or as heterodimers with retinoid X receptor (RXR). Thyroid hormone T3 (3, 3`,5 triiodo-L-thyronine) preferentially binds TR:RXR heterodimers, which activate transcription through coactivator recruitment. However, it is unclear if TR:RXR heterodimers may also be responsive to the canonical RXR agonist 9-cis retinoic acid (9C) in the context of physiological gene regulation. New structural studies suggest that 9C promotes displacement of bound coactivators from the heterodimer, modifying TR:RXR activity. To shed light on the molecular mechanisms that controls TR:RXR function, we utilized biophysical approaches to characterize coregulator recruitment to TR:TR, or to TR:RXR in the presence of T3 and/or 9C, as well as cell-based assays to establish the functional significance of biophysical findings. Using cell-based and fluorescence assays with mutant and wild type TR, we show that 9C does indeed have a function in the TR:RXR heterodimer context, in which it induces the release of corepressors. Further, we show that 9C does not promote detectable conformational changes in the structure of the TR:RXR heterodimer and does not affect coactivator recruitment. Finally, our data supports the view that DNA binding domain and Hinge regions are important to set up NR:coactivator binding interfaces. In summary, we showed that the RXR agonist 9C can regulate TR function through its modulation of corepressor dissociation.