Supplemental Material

This article contains the following supporting material:

• Figure S1 - Effect of Rabex-5 and truncation mutants on EEA1-positive early endosomes. Confocal fluorescence microscopy images showing the morphological changes of GFP-EEA1-labeled early endosomes in BHK cells co-expressing the indicated Rabex-5 constructs. GFP-EEA1 alone (vector) and co-expression with Rab5:Q79L serve as negative and positive controls in determining the enlargement of EEA1-labeled early endosomes. Bar = 16 μm.
• Figure S2 - The MBM is necessary but not sufficient for specific targeting to early endosomes. The confocal fluorescence microscopy images show intracellular localization of the indicated Rabex-5 constructs and co-expressed GFP-Rab5 in BHK cells. The Myc-tagged Rabex-5 constructs are identified by indirect immunofluorescence microscopy with the anti-Myc antibody. Bar = 16 μm. The bottom panel shows membrane and cytosol distribution of Rabex-5(230-399), Rabex-5(135-455), and Rabex-5(48-399) as indicated. Molecular mass standards (in kDa) are indicated on the left.
• Figure S3 - Co-localization of Rabex-5(1-135) with Rab7 on late endosomes. Confocal fluorescence microscopy images showing the localization of Rabex-5(1-135) on late endosomes labeled by RFP-Rab7. The Rabex-5(1-135) construct contains Myc-tag and is identified by immunofluorescence microscopy using the anti-Myc antibody. Bar = 16 μm