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. 2021 Apr 8;11:7730. doi: 10.1038/s41598-021-87259-y

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Subcellular trafficking and melanosome-lysosome segregation of PMEL alanine-scanning mutants. Immunofluorescence analysis of Mel220 cells stably expressing PMEL alanine-scanning mutants. The antibody EP4863(2) recognizes newly synthesized, but not fibrillar PMEL. The PMEL-specific antibody HMB50 recognizes mature melanosomal fibrils. The LAMP1-specific antibody H4A3 recognizes a lysosomal marker. Note that proper PMEL amyloid formation typically correlates in this assay with a significant separation of the perinuclear lysosomal staining (LAMP1) from the largely peripheral melanosomal staining (HMB50).