A previous report demonstrated that nobiletin can induce melanogenesis via up-regulation of extracellular kinase signaling pathways at the later phases; however its ability to induce differentiation in melanoma cells has not been investigated. Nobiletin induced differentiation in murine B16 melanoma cells by increasing tyrosinase activity, melanin content, dendrite formation, and intracellular cAMP, and inhibited cell proliferation in a dose- and time-dependent manners, induced G1 cell cycle arrest, and suppressed phosphorylated protein kinase B (Akt) and extracellular signal-regulated kinase (ERK) protein expressions, which are involved in proliferation. However, nobiletin did not induce DNA fragmentation or condensation indicating potential use of nobiletin to induce differentiation in melanoma cells.