Nuclear receptors are transcription factors regulating the transcription of their target genes in a ligand-dependent manner. They regulate a series of cellular processes including cell growth, differentiation, metabolism, cell death, and immune responses. NRs like retinoic acid receptors (RAR) requires heterodimerization with retinoid X receptors (RXR) for proper transcriptional activity. These receptors diffuse in the nucleus and bind to chromatin even non-specifically or specifically; binding to their hormone response elements. Doxorubicin (Dox) is an anti-cancer drug used as a medicine e.g., for bladder, breast, stomach, soft tissue sarcoma, multiple myeloma cancers. Dox molecules intercalate between DNA bases and thereby inhibit topoisomerase II activity interfering with the transcription process. Dox treatment has many side effects mainly, Dox-induced cardiotoxicity (DIC). Because it rearranges the chromatin, it may interfere with the function of transcription factors. FRAP was used to clarify the impact of dox treatment on the DNA binding properties of RAR. It was observed that dox increased the average mobility of RAR; it decreased the total DNA binding of RAR as indicated by the reduction in τaverage . Interestingly the specific binding of RAR was enhanced in a dose-dependent manner; indicated by the increase in slow fraction and immobile fraction. The reduction of RAR binding induced by Dox may explain the DIC. It could also support the proposal (Dox and retinoic acid) co-treatment strategies applied to augment antitumor and ameliorate side effects.