Figure 1

UL56 shows dynamic localization and relocalizes Nedd4. Images of live cells transiently expressing Nedd4-GFP and/or mRFP-UL56. Time-lapse images were captured with confocal microscopy. (A) Nedd4-GFP is diffusely distributed in the cytoplasm. (B-C') mRFP-UL56 is distributed in a vesicular pattern in the cytoplasm. (C) Images of mRFP-UL56 in the protruding portion of the cell. (C') Six sequential images of the area boxed in C. mRFP-UL56 moves bi-directly; filled arrowheads indicate puncta moving in plus-end direction, and open-arrowheads indicate puncta moving in minus-end direction. (D-F) Nedd4-GFP (shown in green) and mRFP-UL56 (shown in red) colocalizes and partially co-migrated. (E) Magnifications of the perinuclear region. (E-1) Magnification of the area boxed with solid lines in D. (E-2) Six sequential images of the area boxed in E-1. Arrow heads indicate punctuate structures containing both Nedd4-GFP and mRFP-UL56 with low motility (open arrow heads) or high motility (filled arrow heads). (E-3, -4) The punctum indicated with an open arrowhead (E-3) or that indicated with a filled arrowhead (E-4) is magnified. Overlaps between Nedd4-GFP- and mRFP-UL56-localization were not stable. (F) Magnification of the peripheral area boxed with dashed lines in D. Puncta positive for mRFP-UL56 alone and those positive for both mRFP-UL56 and Nedd4-GFP were more abundant in the peripheral of the cytoplasm. Scale bars: 10 μm, in A -E2, F and G; 2 μm, in E3 and E4.