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Example sentences for: gfp-nups
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Three morphological classes include sec mutants with extended networks of ER membrane, those with accumulated Golgi, and a third with accumulated 80-100 nm secretory vesicles [ 37 ] . The morphological perturbations directly correlate with distinct secretory pathway blocks: ER-Golgi, intra-Golgi, and Golgi-plasma membrane [ 44 ] . A recent study in S. pombe has also shown that sec mutants can affect NE and cell cycle progression [ 55 ] . In addition to sec13 , our screen also identified mutant alleles of SEC23 , SEC27 and BET3 . The mislocalization of GFP-Nups in sec13 and sec23 mutants was coincident with the formation of extensive honeycomb networks of ER.
Instead, most mutants lost the exclusive, punctate nuclear rim localization, the GFP-Nups were present in areas distinct from the NE and often in discrete foci (Figure 2, panels G and K).
After 4 hours at 34°, the same conditions that cause GFP-Nup mislocalization in the other assayed sec mutants, the GFP-Nups in sec6-4 remained localized to the NPC in the characteristic, punctate NE rim pattern (Figure 7).
Logarithmically growing cultures of npa1-1/sec23 and npa2-1/sec13 were shifted to the non-permissive temperature as before and then fixed and processed for IF with polyclonal antibodies against Kar2p to visualize the ER [ 45 ] . The monoclonal antibody mAb414, which recognizes a subset of Nups distinct from Nic96p and Nup170p [ 46 ] , was also used because the GFP fluorescence of the GFP-Nups was more sensitive to fixation when the GFP-Nups were mislocalized than when they remained at the nuclear rim (unpublished observation).
In comparison to our previous genetic screen with a strain expressing only GFP-Nup49p [ 26 ] , these results suggest that increasing the number of GFP-Nups as reporters for NPC integrity effectively increases the likelihood of obtaining a panel of mutants defective in global NPC assembly, stability, and localization.