Distinct Protein Sorting and Localization to Premelanosomes, Melanosomes, and Lysosomes in Pigmented Melanocytic Cells -- Data Supplement - JCB -- Raposo et al. 152(4): 809 Figure S3 -- The Journal of Cell Biology

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Figure S3.The coated endosome is an intermediate in the targeting of Pmel17 to the stage II premelanosome. Double immunogold labeling with two distinct antibodies directed against Pmel17. ${alpha}$ PEP13h is directed to the cytoplasmic domain of Pmel17, which becomes inaccessible within the stage II premelanosome. HMB45 is directed to the lumenal domain of Pmel17; recognition by HMB45 requires a conformational change or processing event that occurs in the Golgi apparatus or TGN. (A) Both HMB45 (lum PAG 5) and ${alpha}$ PEP13h (Cyt-PAG 10) recognize Pmel17 in the coated endosome (CE). (B) The ${alpha}$ PEP13h antibody (cyt -PAG 10) recognizes Pmel17 in the Golgi appara-tus (GA) and small closely apposed vesicles, whereas HMB45 (lum-PAG 5) only reacts poorly with Pmel17 in these compartments. In contrast, stage II melanosomes (II) are highly labeled with HMB45, but no labeling is observed with ${alpha}$ PEP13h. Bars, 100 nm.