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Figure 9


Fig. 9. UAP56 K95N has a dominant-negative effect on RNA export. 293T cells were co-transfected with a β-globin-splicing reporter plasmid and EGFP-UAP56 wt or EGFP-UAP56 K95N expression vector. 24 hours after transfection, the RNA of the cells was separated into cytoplasmic and nuclear fractions. Real-time PCR with primers recognizing the 5' exon and 3' exon was performed. (A) In cells expressing EGFP-UAP56 K95N, there was a 15-fold increase of β-globin in the nuclear fraction compared with cells expressing EGFP-UAP56 wt. (B) When the nuclear PCR product was separated on a 1% agarose gel it was all spliced, showing that the nuclear accumulation is due to reduced mRNA export and not pre-mRNA splicing.





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