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. 2009 Dec 15;285(8):5941–5953. doi: 10.1074/jbc.M109.039925

FIGURE 3.

FIGURE 3.

The region of p62 between amino acids 303 and 320 is sufficient for nuclear export. A, the nuclear export activity of p62 region 303–320 was tested using the pRev1.4-GFP assay system where putative NESs were inserted between the nuclear export-impaired Rev protein from HIV and GFP. B, representative images are shown of HeLa cells transiently transfected with Rev1.4-GFP, Rev1.4-NES-GFP, or Rev1.4-p62-(303–320)-GFP vectors (upper panel). Nucleocytoplasmic shuttling of Rev1.4-p62-(303–320)-GFP was verified by LMB treatment (lower panel). Scale bars equal 10 μm C, quantification of nuclear/cytosolic distribution of transiently transfected constructs from panel B is shown. Labels are the same as in Fig. 2E. Each bar represents the mean ± S.D. of 3 independent experiments with more than 100 cells counted per experiment. D, overexpression of the NES from Rev or the p62 fragment 303–320 inhibits the nuclear export of endogenous (end.) p62. Representative are shown of images of HeLa cells transiently transfected with indicated constructs, stained with anti-p62 polyclonal antibody. Scale bars equal 10 μm.