The steroid hormones estrogens have an influence on many physiological and developmental processes in vertebrates. The estrogen receptor α (ERα), which is a ligand-regulated transcription factor, is one of the effectors of estrogens. Besides hormonal signals, the regulation of ERα function is mediated by a large number of factors acting at different steps in the ERα signaling pathway. We sought to define the degree of dependence of ERα activity toward the nuclear transport receptor Exportin-5 (Exp-5), which exports various RNA and protein species out of the nucleus. Our interest was oriented on Exp-5 since previous work from our laboratory suggested that in the budding yeast the Exp-5 ortholog Msn5p was essential for proper function of the human ERα. We used different experimental strategies that were principally based on the depletion of Exp-5 coupled with assays monitoring the activity of ERα in tissue culture cells. In addition, we evaluated the impact of Exp-5 knockdown on the subcellular localization of ERα. Finally, we employed an in vitro approach to determine whether these two proteins physically interact. In this context, we also used bioinformatics in order to identify proteins that interact both with ERα and with Exp-5. Overall, although we did not collect solid enough data to draw firm conclusions, we obtained several results hinting at the involvement of Exp-5 in the regulation of ERα activity.