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. 1999 Mar 22;144(6):1097–1112. doi: 10.1083/jcb.144.6.1097

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Proteolytic processing of the Nup98-Nup96 precursor is essential for targeting Nup98 and Nup96 to the NPC. HeLa cells were transiently transfected with: (A–C) the wild-type Nup98-Nup96 precursor, (D–F) the mutant Nup98-Nup96 precursor (F863S/Y866R), (G–I) the Nup98 domain (amino acids 1–863), and (J–L) Nup96 (amino acids 864–1712). Localization of the transiently expressed proteins was detected by indirect immunofluorescence and confocal microscopy using an anti-myc epitope antibody followed by a Cy3-conjugated secondary antibody (A, D, G, and J). Cells were also double labeled with an antibody specific for Nup358, which was detected using a secondary antibody conjugated to FITC (B, E, H, and K). Cy3 and FITC images were merged to identify coincident labeling of NPCs (C, F, I, and L). Bar, 10 μm.