Supplemental Material

This article contains the following supporting material:

• Figure 1 - Immortal melan-ln melanocytes closely resemble primary cultured ln melanocytes. Panels A-C. Lysates were prepared and PNS proteins (50μg) subjected to SDS-PAGE followed by immunoblotting using specific antibodies to detect Mlph (A), Rab27a (B) and MyoVa (C) (as described under Materials and Methods). Panels D-M. Cover-slip grown melan-ln melanocytes were fixed 48hrs after transfection and stained using the indicated antibodies (as described under Materials and Methods). In panels D-K cells were transfected with pCS26xMycMlph as a control to show the specificity of the Mlph antibodies (D-G) and to highlight the differences in distribution of MyoVa in the presence and absence of exogenous Mlph in melan-ln cells (H-K). Panels D and H show the pattern of over-expressed myc tagged Mlph. Panels E, I and L show the distribution of the known components of the melanosome transport complex (E-Mlph, I-MyoVa and L-Rab27a). Panels F and J show merged fluorescence images (green = anti-Mlph or MyoVa, red = anti-myc). Panels G, K and M show the corresponding transmission images. Bar = 20μm.