The Nsp1 complex has a cytoplasmic pool localized to the daughter bud. (A) Fluorescence micrographs (and bright field) of Nsp1-GFP– and Nup170-GFP–expressing cells (CPL1234, BWCPL437, PCCPL423, and PCCPL409) arrested in G1, S, or G2 phase with α-factor–, hydroxyurea-, or galactose-induced overexpression of SWE1, respectively. To visualize cytoplasmic foci, we show maximum-intensity projections of a deconvolved z series where 0.4% of pixels have been digitally saturated. Bars, 5 µm. (B) Plot showing the percentage of cells where the indicated nups were found in cytoplasmic foci in α-factor– or hydroxyurea-arrested cells. 90 ≤ n ≤ 190. Error bars indicate standard error of the mean. (C) Plot showing the number of Nsp1-GFP_CYT foci and their distribution in daughter buds and mother cells during 3 h of a G2 arrest. At the indicated times, >100 cells were scored. Error bars indicate standard deviations from the mean. Bud size is the surface area of the bud (µm²). (D) NSP1-(PCCPL506)– and NUP85-RITE (PCCPL523)–containing strains were grown in the presence of galactose (to induce a G2 arrest) and estradiol and imaged over several hours (see Videos 2 and 3). Fluorescence micrographs (maximum-intensity projections of a deconvolved z series) of the GFP and mCherry forms of Nsp1 and Nup85 are shown at the indicated time points. Note the specific appearance of Nsp1-GFP “new” (arrowheads) in the elongated bud. Bars, 5 µm.