NPC association of activated INO1 requires the Scs2–Siz2 complex and Scs2 SUMOylation. NE localization of the activated INO1 locus was examined using epifluorescence imaging. The graphs show the percentage of total GFP-lac/INO1-LacO₂₅₆ foci that colocalize with NE localized Nup49-RFP. For each indicated strain, three biological replicates were assessed. Cell cycle stage was determined using bud size and nuclear morphology. n = 50 cells/replicate/cell cycle stage. Error bars represent SD. Asterisks: significant change relative to WT using a two-tailed Student’s t test. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001.