Figure 4.

Assessment of the role of various karyopherins on nuclear import of over‐expressed Aim44 and Nis1. (a) a CEN plasmid expressing Aim44‐mNG under control of the GAL1/10 promoter was transformed into WT cells and otherwise isogenic importin knockout strains pse1Δ, mtr10Δ, nmd5Δ or kap123Δ. Cells were grown in a medium containing galactose to induce expression of Aim44‐mNG for 3 h prior to imaging. (b) a CEN plasmid expressing Nis1‐mNG under control of the GAL1/10 promoter was transformed into WT cells and otherwise isogenic importin knockout strains pse1Δ, mtr10Δ, nmd5Δ or kap123Δ. Cells were grown as in (a) to induce expression of Nis1‐mNG. (c) Quantitation of the experiment depicted in panel (a). Gray bars, percentage of cells displaying Aim44‐mNG nuclear localization for each strain; white bars, ratio of nuclear‐to‐cytoplasmic pixel intensity for those cells that displayed a fluorescent signal at the nucleus. Values represent the average from three independent experiments, in each of which ≥50 cells were examined; error bars represent standard error of the mean. Double asterisks (**), p < .05; triple asterisks (***), p < .001, as determined by a two‐tailed Student's t‐test. (d) Quantitation of the experiment depicted in panel (b), performed as described in panel (c). scale bars = 1 μm