Lagging chromosomes that form micronuclei farther from the spindle pole are more likely to experience micronuclear envelope rupture. (A) Plot of locations where micronuclei formed relative to the pole after nocodazole washout as a function of time of micronuclear envelope rupture in RPE1 cells stably expressing Centrin-1-GFP and CENP-A-GFP and transfected with mCherry-H2B and NLS-EGFP. n = 31 micronuclei (17 remain intact and 14 rupture) collected across 14 independent experiments. Dashed line represents the average measured position of micronucleus formation for micronuclei that experienced micronuclear envelope rupture: 7.7 ± 1.2 μm from closest spindle pole. (B) Representative still images of a cell with two lagging chromosomes. Lagging chromosome farther from the pole is indicated by a white arrowhead. Bottom panels are annotated to indicate position of Centrin foci (green circles) and the distance between farthest lagging centromere and the pole (yellow line). (C) Still frames of daughter cells from division shown in B. Micronucleus that formed around lagging chromosome farthest from the pole indicated by white arrowhead in top panels. Micronuclear envelope rupture denoted by loss of micronuclear NLS-EGFP signal (green) from micronuclear chromatin (mCherry-H2B, red) is indicated by red arrowhead in bottom panels.