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. 2017 Mar;23(3):365–377. doi: 10.1261/rna.057612.116

FIGURE 2.

FIGURE 2.

tRNAs are enriched in the nuclei of cells specifically lacking the GLFG domain of Nup100. (A–C) (Top) Representative images of fixed cells from the listed W303 strains incubated with Cy5-labeled probes that hybridize tRNAile (A), tRNAtrp (B), or tRNAtyr (C). DAPI was used to stain DNA and is shown below each Cy5 image. Scale bar, 5 µm. (Bottom) Nuclear:cytoplasmic Cy5 ratios of the cells shown above were determined; each gray dot represents the ratio from one cell, and 60 cells were quantified for each strain (20 cells from three separate experiments). Large lines represent the averages for each strain, while error bars represent the 95% CI. Averages are listed below dots. An asterisk indicates a P-value of less than 0.05 when compared to wild type using Tukey's post-hoc test following a one-way ANOVA. (D,E) (Top) Representative images of S288C nup100Δ cells transformed with pRS313, pRS313-NUP100, or pRS313-nup100ΔGLFG and incubated with Cy5-labeled probes that hybridize tRNAtyr (D) or tRNAtrp (E). DAPI was used to stain DNA and is shown below each Cy5 image. Scale bar, 5 µm. (Bottom) Nuclear:cytoplasmic ratios for cells shown above were calculated; an asterisk indicates a P-value of less than 0.05 when compared to nup100Δ cells transformed with pRS313 using Tukey's post-hoc test following a one-way ANOVA with at least 30 cells.