FIGURE 5.

Disruption of Nup50-Nup153 interaction prevents nuclear rim localization of Nup50. A, HeLa cells were engineered to express either GFP or GFP-Nup153(401–609) in response to doxycycline (DOX). Cells that were either untreated or incubated with doxycycline for 24 h were harvested, and the cell lysates were probed for levels of Nup50, Tpr, and Nup153 as indicated. The induction was confirmed by tracking GFP fusion proteins, and α-tubulin (αTub) was tracked to ensure equivalent loading of samples. B, following doxycycline-induced expression of GFP (left panels) or before (middle panels) and after (right panels) similarly induced GFP-Nup153(401–609) expression, the localization of Nup50 was tracked by indirect immunofluorescence, with DNA detection by DAPI staining shown in the accompanying panels. C, the nucleoporins Nup153 and Tpr, as well as lamin, were detected by indirect immunofluorescence under similar conditions in which either GFP or GFP-Nup153(401–609) had been induced by doxycycline. Scale bars = 10 μm.