PNU-74654 is primarily utilized in research settings, particularly in the study of Wnt signaling pathways. It functions as an inhibitor of β-catenin/TCF interactions, which are crucial components of the Wnt signaling cascade. By disrupting this interaction, PNU-74654 helps researchers investigate the role of Wnt signaling in various biological processes, including cell proliferation, differentiation, and cancer development. Its application is significant in understanding the molecular mechanisms of diseases where Wnt signaling is dysregulated, such as certain types of cancers. Additionally, it serves as a valuable tool in drug discovery efforts aimed at developing therapeutic agents targeting Wnt-related pathologies.