Fig. 4: Reconstitution of B55α in PC3 and DU145 iB55α cells blocks mitotic exit as a result of extended mitotic checkpoint and chromosome segregation failure. | Oncogenesis

Fig. 4: Reconstitution of B55α in PC3 and DU145 iB55α cells blocks mitotic exit as a result of extended mitotic checkpoint and chromosome segregation failure.

From: PPP2R2A prostate cancer haploinsufficiency is associated with worse prognosis and a high vulnerability to B55α/PP2A reconstitution that triggers centrosome destabilization

Fig. 4

a, b PC3 iB55α cells were synchronized by Nocodazole, shaken off, and reseeded as scheme shown. a B55α reconstitution slowed cell cycle progression through G2/M. b Expression of F-B55α, B55α, cyclin B1 and the indicated proteins was determined via western blot analysis. c, d Mitotic defects resulting from B55α reconstitution in DU145-iB55α-EGFP-H2B cells were determined by live imaging using confocal microscopy. Three fields of each treatment were imaged in at least three independent experiments. c Untreated cells went through normal mitosis in about 60 min. d Dox induced B55α caused an extended metaphase checkpoint followed by chromosome segregation failure and apoptosis. e Co-expression of EGFP-H2B and mRFP-α-tubulin in DU145-iB55α cells allowed visualization of bipolar spindles prior to centrosome collapse when B55α is reconstituted. Lens: ×20. f DU145-iB55α cells were synchronized by nocodazole as in a, reseeded and collected at the indicated time points. Cell cycle analysis was determined via PI/FACS in at least two independent experiments.

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