Localization of NPCs in proximity to duplicating SPBs. (A) Immuno-EM analysis with α-Nsp1 antibody of SPC42 SPC29 OE cells with 45 min induction during α-factor arrest and after release into the cell cycle. EM enlargements on the right show NPCs labeled by the α-Nsp1 antibody from the NE of the “inserted fusion SPB” cell. Cartoons illustrate SPB phenotypes and gold particles (pink). B, bridge; N, nucleus; nMT, nuclear MT; S, satellite. Bars: (main images) 200 nm; (insets) 50 nm. (B) Phenotype analysis of EM images (Figs. 1, 2, and 3). n > 100 for the entire dataset. (C) Quantification of EM micrographs categorized according to their phenotypes and analyzed for the distance between the mSPB/satellite and NPCs. In the three bottom categories, the G1 block was released. n, number of analyzed SPBs. Red-encircled numbers indicate NPCs (%) at the distal end of the bridge. (D) Representative images and quantification of SIM analysis in SPC42-yeGFP NIC96-tdTomato WT cells in α-factor arrest and upon release. n ≥ 30. Bars: (main images) 2 µm; (insets) 500 nm.