Quantitative analyses were performed using the Object Count tool in Nikon AR 4.60. The entire cell was imaged to ensure that no specific region of the cell was favored; however, when analyzing each cell, an equivalent diameter (EqDiameter) was restricted to 1.85 to 15.00 pixels and circularity to 0.20 to 1.00 for optimal identification of individual puncta and exclusion of larger structures such as the Golgi apparatus. The lower intensity threshold limit of each fluorescence channel was defined as the intensity of the dimmest punctum returned using the 3 points circle threshold tool. The upper intensity threshold limit was set to the maximum value. This method assured that while the intensity of fluorescence changed between cell lines and conditions, all melanosome specific data were captured and subjected to analysis.

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