Hair cells (HC) are mechanosensory receptors that reside in the inner ear and are responsible for hearing in vertebrates. While damage to the mammalian inner ear HCs is irreversible, non-mammalian species have an innate capacity for hair cell regeneration (HCR) after injury to HCs. In zebrafish, regenerative capacity of HCs depends on the activation of a pool of surrounding cells called supporting cells (SCs) into new HCs, but the mechanistic details of the pathway controlling this transition remain elusive. Previous single-cell experiments suggest that during HCR, the SOX transcription factors (TF) are recruited to enhancers in the supporting cells to trigger a regenerative response, while both SOX and SIX family of transcription factors are recruited to enhancers in cell types that have already been specified to lock in a HCs specific fate. Of particular interest is the SOX2 TF, whose expression increases significantly as SC transdifferentiate, and altering expression of SOX2 causes a disruption in HCR in the inner ear. Considering SOX2’s known role as a stem cell pluripotency factor that can interact with regulatory proteins in complexes such as SOX5/6 and OCT4 during development, we are positioned to address if SOX2 is able to interact with other proteins in zebrafish to successfully drive the HCR program. We hypothesize that SOX2 is involved in binding interactions with proteins that co-regulate DNA expression and impact SOX2’s ability to drive HCR. We describe a method to purify SOX2 complexes from larvae whole-body cells and adult zebrafish inner ear HCs. We have successfully generated a zebrafish line with the HA (hemagglutinin) tag attached to SOX2 and detected intracellular SOX2-HA complexes with an anti-HA tag antibody. Co-immunoprecipitation of larval zebrafish cells containing SOX2-HA complexes has revealed at least one major SOX2 complex in larval zebrafish and will now be confirmed using mass spectrometry.