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. 2006 Jun 19;173(6):917–926. doi: 10.1083/jcb.200510010

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Copyright © 2006, The Rockefeller University Press

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Figure 6. — TIP47 binding to Rab5/9 triggers relocalization from early to late endosomes. (A) Wide-field light microscopy. GFP-Rab5/9 localization in fixed HeLa cells (left). The overexpression of wild-type TIP47 (middle), but not of mutant TIP47^SVV-AAA (Hanna et al., 2002), which is deficient for Rab9 binding (right), causes GFP-Rab5/9 localization to perinuclear late endosomes. (middle) The cell is outlined in white. Levels of TIP47 wild-type and mutant protein overexpression were approximately ninefold. (B) TIP47-relocalized perinuclear Rab5/9 is present on late endosomes. Deconvolution microscopy of cells overexpressing TIP47 protein; perinuclear GFP-Rab5/9 colocalized with late endosomal, endocytosed anti–CI-MPR antibody. (C) Deconvolution microscopy shows TIP47- relocalized perinuclear Rab5/9 is not present on the Golgi, as it did not colocalize with the Golgi marker p115. In the merge images in B and C, Rab5/9 is shown in green, whereas the organelle markers are shown in red. In B and C, the TIP47 is shown as the mean of the summation of total Z-sections; other images are 0.2-μm Z-sections. Bars: (A) 10 μm; (B and C) 5 μm.