Files in this Data Supplement:
- Supplemental Materials (PDF)
- Video 6 -
Behavior of nuclear GLT1 mRNAs in a Δmlp1/2 strain after galactose induction. Diploid WT yeast cells containing a dTomato-tagged Nup188 (red) and where 24xPP7 stem loops were inserted to the 5′ UTR of one allele of the galactose-inducible GLT1 reporter gene. GLT1 mRNAs were visualized by transforming a plasmid expressing PP7-2xGFP (green) and inducing transcription by galactose. mRNAs were analyzed during the early time of induction. Images were acquired using a spinning disk confocal microscope, and frames were taken every 37 ms. The video is played at 10 frames per second. - Video 2 -
Nuclear scanning of CLB2 mRNA occurs outside of the nucleolus. Diploid WT yeast cells where 12xPP7 stem loops were inserted into the 3′ UTR of one allele of the CLB2 gene, and the nucleopore protein Nup188 and the nucleolar marker Gar1p were C-terminally tagged with mCherry (red). CLB2 mRNAs were visualized by transforming a plasmid expressing PP7-2xGFP (green). Images were acquired using a spinning disk confocal microscope, and frames were taken every 37 ms. The video is played at 10 frames per second. - Video 1 -
Nuclear scanning of CLB2 mRNA. Diploid WT yeast cells where 12xPP7 stem loops were inserted into the 3′ UTR of one allele of the CLB2 gene and the nucleopore protein Nup188 was C-terminally tagged with mCherry (red). CLB2 mRNAs were visualized by transforming a plasmid expressing PP7-2xGFP (green). Images were acquired using a spinning disk confocal microscope (Observer; Carl Zeiss), and frames were taken every 37 ms. The video is played at 10 frames per second. - Tables S2 and S3 (zipped Excel files) -
Table S2 summarizes the proteins identified by mass spectrometry purified using Mlp1-ProtA or Mlp1ΔC-ProtA as baits (Fig. 4, B and C). Table S3 lists the smFISH probes. - Video 5 -
Nuclear scanning of galactose-induced GLT1 mRNA. Diploid WT yeast cells containing a dTomato-tagged Nup188 (red) and where 24xPP7 stem loops were inserted to the 5′ UTR of one allele of the galactose-inducible GLT1 reporter gene. GLT1 mRNAs were visualized by transforming a plasmid expressing PP7-2xGFP (green) and inducing transcription by galactose. mRNAs were analyzed during the early time of induction. Images were acquired using a spinning disk confocal microscope, and frames were taken every 37 ms. The video is played at 10 frames per second. - Video 7 -
Behavior of nuclear GLT1 mRNAs in a Δtom1 strain after galactose induction. Haploid WT yeast cells containing a dTomato-tagged Nup188 (red) and where 24xPP7 stem loops were inserted to the 5′ UTR of the galactose-inducible GLT1 reporter gene. GLT1 mRNAs were visualized by transforming a plasmid expressing PP7-2xGFP (green) and inducing transcription by galactose. mRNAs were analyzed during the early time of induction. Images were acquired using a spinning disk confocal microscope, and frames were taken every 37 ms. The video is played at 10 frames per second. - Video 3 -
Nuclear scanning of MDN1 mRNA occurs outside of the nucleolus. Diploid WT yeast cells where 12xPP7 stem loops were inserted into the 3′ UTR of one allele of the MDN1 gene, and the nucleopore protein Nup188 and the nucleolar protein Gar1p were C-terminally tagged with mCherry (red). MDN1 mRNAs were visualized by transforming a plasmid expressing PP7-2xGFP (green). Images were acquired using a spinning disk confocal microscope, and frames were taken every 37 ms. The video is played at 10 frames per second. - Video 4 -
MDN1 mRNA trapped and exported through the nucleolus. Diploid WT yeast cells where 12xPP7 stem loops were inserted into the 3′ UTR of one allele of the MDN1 gene, and the nucleopore protein Nup188 and the nucleolar protein Gar1p were C-terminally tagged with mCherry (red). MDN1 mRNAs were visualized by transforming a plasmid expressing PP7-2xGFP (green). Images were acquired using a spinning disk confocal microscope, and frames were taken every 37 ms. The video is played at 10 frames per second.