We recently reported that the oral intake of β-cryptoxanthin exerted anti-obesity effects by lowering visceral fat levels. In the present study, we characterized the molecular mechanisms underlying the lipid-lowering effects of β-cryptoxanthin on 3T3-L1 cells. Consistent with our previous findings, β-cryptoxanthin rapidly reduced the level of intracellular lipids in 3T3-L1 cells as assessed by Oil red O staining. Using an in vitro nuclear receptor binding assay, we demonstrated the ability of β-cryptoxanthin to bind to and activate members of the retinoic acid receptor (RAR) family. Accordingly, treatment of cells with LE540, an RAR antagonist, abolished the β-cryptoxanthin-dependent suppression of 3T3-L1 adipogenesis, suggesting that β-cryptoxanthin mediates its effects on 3T3-L1 cells via RAR activation. In addition, real-time RT-PCR analysis revealed that β-cryptoxanthin down-regulates mRNA expression of PPARγ, a key regulator of adipocyte differentiation, and that this inhibition was blocked by LE540 treatment. Taken together, these data indicate that RAR activation contributes to the molecular mechanism by which β-cryptoxanthin prevents obesity.