Fig. 2: Inactivation of CIP2A disrupts mitotic clustering and disperses pulverized chromosome fragments.
From: Mitotic clustering of pulverized chromosomes from micronuclei
a, DOX/IAA-treated mitotic DLD-1 cells with the indicated genotypes with γH2AX-positive Y chromosomes. sg3 and sg4 denote distinct sgRNAs targeting exon 1 of CIP2A. b, Fragment clustering from γH2AX-negative and γH2AX-positive Y chromosomes from a. Data are mean ± s.e.m. Statistical analysis was performed using one-way ANOVA with correction for multiple comparisons compared with the WT controls; ****P ≤ 0.0001. n = 3 independent experiments; from top to bottom, 657, 455, 352, 526, 383, 238, 165, 175, 179 and 149 cells. c, Mitotic CIP2A-KO (sg3) cell with γH2AX-positive Y-chromosome fragments displaced from the genomic mass (arrows). d, Fragment clustering from γH2AX-positive Y chromosomes from CIP2A-KO cells rescued by ectopic CIP2A–HaloTag. Data are mean ± s.e.m. Statistical analysis was performed using two-tailed t-tests compared with CIP2A-KO cells; ****P ≤ 0.0001, ***P = 0.0002. n = 3 independent experiments; 137 (WT), 200 (no rescue), 139 (full length (FL)) and 115 (ΔNES) cells. e, Schematic of inducing the depletion of CIP2A fused to a FKBP12(F36V) degron with dTAGv-1 in mitosis-synchronized cells. noc., nocodazole. f, Immunoblot analysis showing rapid degradation of CIP2A–FKBP12(F36V) after 4 h treatment of mitotic cells with dTAGv-1. Async, asynchronous; sync., synchronized. g, Clustered and dispersed Y-chromosome fragments after mitotic CIP2A depletion. h, Fragment clustering from γH2AX-positive Y chromosomes from g. Data are mean ± s.e.m. Significance was determined using two-tailed Student’s t-tests. n = 3 independent experiments; 194 (mock) and 188 (dTAGv-1) cells. For a,c and g, scale bars, 5 μm.
