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In this study, we showed that in melanocytes derived from transgenic mice expressing active β-catenin (bcat* cells), pigmented melanosomes were primarily absent from the perinuclear area and localized at the cell periphery. Notably, the expression of Dynlt3, a member of the cytoplasmic dynein complex, was downregulated in bcat* cells, and knockdown of Dynlt3 in WT melanocytes phenocopied the peripheral localization of pigmented melanosomes and their absence from the perinuclear area. Melanosome movement was increased and melanosomes moved in a more directional manner in cells with diminished levels of Dynlt3. Decreased levels of Dynlt3 resulted in more acidic melanosomes that were associated with a reduction of melanosome transfer to keratinocytes. Altogether, these results show that Dynlt3 is a key player in regulating melanosome movement, acidity, and transfer. As a target of β-catenin, our results reveal a function for the Wnt/β-catenin signaling pathway during melanocyte and skin pigmentation. 2b1af7f3a8