Extended Data Fig. 3: Characterization of human DLD-1 cells harbouring biallelic deletions in CIP2A.
From: Mitotic clustering of pulverized chromosomes from micronuclei
a) Immunoblot confirmation of CIP2A KO clones. b) Growth curves of WT and CIP2A KO cells with and without DOX/IAA treatment over the indicated number of days. Data represent mean ± SEM; n = 3 biological replicates. c) Flow cytometry analysis of propidium iodide-stained WT and CIP2A KO cells showing similar cell cycle distribution profiles. d) Proportion of micronucleated cells with or without 2d DOX/IAA induction, as determined by DAPI staining. Data represent mean ± SEM; WT: **P = 0.0017, sg3: **P = 0.0023, sg4: *P = 0.0261 by two-tailed t-test compared to untreated controls; n = 3 independent experiments; WT (-DOX/IAA = 5,521, +DOX/IAA = 3,718), CIP2A KO sg3 (-DOX/IAA = 3,436, +DOX/IAA = 2,450), CIP2A KO sg4 (-DOX/IAA = 3,999, +DOX/IAA = 2,930 cells). e) Frequency of Y chromosome fragmentation among Y chromosome-positive metaphase spreads following 3d DOX/IAA induction. Data represent mean ± SEM; not significant (ns), P > 0.05 by two-tailed t-test compared to WT controls; n = 3 independent experiments; WT = 234, CIP2A KO sg3 = 269, CIP2A KO sg4 = 284 cells. f) Mitotic CIP2A KO cells exhibiting fragment dispersion with and without cell cycle arrest with the indicated mitotic inhibitors. Dispersion frequencies and the number of cells analysed are shown. g) Immunoblot of ectopic CIP2A-HaloTag complementation in CIP2A KO cells generated by a frameshift deletion in exon 3 induced by Cas9 ribonucleoprotein (sgRNP) delivery. FL, full length; NES, nuclear export signal.
