tra12136-sup-0001-FigureS1.docWord document27KFigure S1: Quantification of peri-nuclear and cytoplasmic protein. To quantify perinuclear/cytoplasmic localization, a macro was created in ImageJ* (version 1.47i). The macro is fully automated with the only user interaction required being the selection of the directory containing the images to be quantified. Two channels are utilized by the script from each image: a DAPI channel to define the nuclear regions (blue channel); and the protein of interest (POI) channel (green channel). For each image, the ratio of total protein found in the perinuclear to cytoplasmic regions (as defined by distance from the nuclei in the images) is then calculated and recorded in an Excel file. See Materials and Methods for details of the quantification process.
tra12136-sup-0002-AppendixS1.avivideo/avi1580KMovie S1: Vps35 WT-GFP positive endosomes are mobile and found throughout the cytoplasm. Details in Figure legend 2A.
tra12136-sup-0003-AppendixS2.avivideo/avi1081KMovie S2: Vps35 D620N-GFP positive endosomes are enlarged and redistributed to a perinuclear localization but retain ability to receive endocytosed material. Details in Figure legend 2A.

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