The distribution of Nsp1_CYT and the density of daughter NPCs require MYO2. (A) Maximum-intensity projections of a deconvolved z series of images showing the distribution of Nsp1-GFP in wild-type (WT; CPL1234), myo2-14 (PCCPL317), myo2-20 (PCCPL316), dyn1Δ (PCCPL559), dyn2Δ (PCCPL427), and myo4Δ (PCCPL365) cells after 3 h of arrest in hydroxyurea at RT. Arrowheads point to Nsp1-GFP_CYT foci. Bars, 2 µm. (B) The number and distribution of Nsp1-GFP_CYT foci were assessed in the indicated hydroxyurea-arrested cells. For each cell it was determined whether there was a bud-biased (BB), unbiased (UB), or mother-biased (MB) localization in the total number of Nsp1-GFP_CYT foci per cell. These numbers were plotted as a percentage of total cells. 40 ≤ n ≤ 60. Error bars are standard deviations from the mean. (C) Deconvolved fluorescence micrographs showing a middle z section of WT (BWCPL42), myo2-14 (PCCPL529), dyn1Δ (PCCPL561), dyn2Δ (PCCPL445), and myo4Δ (PCCPL530) cells expressing Nup85-GFP after anaphase. The bottom panel is a “heatmap” representation of fluorescence intensities normalized to a 1-256 arbitrary scale (legend on the left) of the same mother (M) and daughter (D) cells as in the top panel. Cell boundaries are denoted by outlines. Bars, 2 µm. (D) In a middle z plane of a deconvolved z series, the mfi of the NE of a mother and daughter cell were measured and plotted as a ratio (mfi_d/mfi_m) as an indirect readout of relative NPC density. 50 ≤ n ≤ 98. Box plot and statistics are as in Fig. 3. P < 0.0001.