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. 2014 Oct 22;289(49):33767–33782. doi: 10.1074/jbc.M114.616466

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© 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 4. — C-terminal region of FLASH targets SLBP and GFP to HLBs in HeLa cells. A, HA-tagged versions of SLBP (HA-SLBP) or SLBP extended at the C terminus by the last 103 amino acids of FLASH (HA-SLBP/F) were transiently expressed in HeLa cells and their levels compared relative to the endogenous SLBP using an anti-SLBP antibody (αSLBP). B, HeLa cells transiently expressing HA-SLBP (top panels) and HA-SLBP/F (bottom panels) were assayed by immunofluorescence. HLBs were detected by the αFLASH/N antibody and stained green. The intracellular localization of the HA-SLBP and HA-SLBP/F fusion proteins was detected using a mouse anti-HA antibody (αHA) and stained red. Nuclei were visualized with DAPI. HA-SLBP/F, but not HA-SLBP, in addition to showing a diffuse nuclear localization is enriched in a number of distinct foci that co-localize with the HLBs detected by the αFLASH/N antibody. C, immunofluorescence of HeLa cells transiently expressing GFP/F+NLS (top panels) and GFP/F (bottom panels). HLBs were detected using a rabbit antibody against N-terminal FLASH (αFLASH/N) and stained red. Nuclei were visualized with DAPI.