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Fig. 4. Drosophila Nup214 attenuates CRM1-mediated protein export. (A) GFP-NES was expressed under the control of the hsp70 promoter in wild-type, emb³ and nup214 mutant larvae and detected with a GFP antibody (red). All panels show larval gut cells. Nuclei were visualized by DAPI staining (middle panels). Bar, 35 µm. (B) Nuclear export rates of GFP-NES are enhanced in nup214 mutants. The ratios of nuclear:cytoplasmic GFP-NES intensities of early second (eL2) and early third (eL3) instar wild-type, nup214 and emb³ larval gut cells are shown in a log₂ graph. The nuclear accumulation of the GFP-NES reporter is decreased 30% in nup214 mutants, whereas in emb³ mutants the nuclear accumulation is increased by 40% (P<0.0001 by pair-wise t-test). Error bars represent s.e.m. (C) Confocal sections of fat body cells from wild-type and nup214 mutant larvae stained with anti-CRM1, anti-lamin and anti-RanGAP antibodies. Error bars, 5 µm. (D) Western blot of protein extract from wild-type and nup214 mutant larvae probed with anti-CRM1. ß-tubulin provides a loading control. (E) Quantification of RanGAP levels along the nuclear rim in wild-type and nup214 mutants. The nuclear rim staining in the mutants is reduced by 35%.

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