The mechanisms distinguishing successful cardiac regeneration from fibrotic healing remain poorly understood. In response to myocardial infarction, adult mammals typically initiate an inflammatory cascade that culminates in fibrosis, whereas neonatal mice exhibit the unique ability to regenerate cardiomyocytes (CMs). Our study employed comparative spatiotemporal single-cell analysis and genetic tools to explore the role of Clusterin (CLU) in heart regeneration. To identify CLU’s target proteins, we conducted immunoprecipitation to enrich CLU, followed by mass spectrometry to characterize its interacting proteins.