Cdk4 6i palbociclib

Cells were seeded to 60–70% density. For mitotic slippage induction, 0.1 μg ml^–1 nocodazole (Sigma-Aldrich, M1404) was added to the growth medium and CP-A and K562 cells were incubated for 72 and 48 h, respectively. For cells with an elongated G1 phase after tetraploidization, WGD in  CP-A TP53^−/− cells was induced with 0.1 μg ml^–1 nocodazole for 72 h and treated with 0.5 μM of the CDK4/6i palbociclib (Sigma-Aldrich, PZ0383) for the last 16 h of the WGD induction protocol. For cytokinesis failure inductions, RPE cells were incubated for 24 h with 0.1 μg ml^–1 nocodazole-containing medium. Following nocodazole treatment, the cells were exposed for an additional 24 h to 4 μM dihydrocytochalasin B (Cayman Chemical, 20845). The treatment was removed and cells were allowed to recover for 48 h to allow transition from a binucleated to a mononucleated state. Alternatively, WGD was induced through cytokinesis failure in RPE TP53^−/− cells by incubation for 24 h with 9 μM of the CDK1 inhibitor RO-3306 (Sigma-Aldrich, SML0569) for G2 synchronization. The compound was washed off and cells were then treated with 4 μM dihydrocytochalasin B for 24 h. Cells were allowed to recover for 48 h, and tetraploid cells were sorted on the basis of cell cycle staining with 1 μg ml^–1 Hoechst 33342 (Thermo Fisher Scientific, H1399).

β-estradiol (E2), fulvestrant (ICI182;780; Tocris) and testosterone (Sigma-Aldrich) were dissolved in ethanol. CDK2is dinaciclib (SCH727965; S2768, Selleckchem), SNS-032 (S1145, Selleckchem), PF-06873600 (A16875, Adooq) and fadraciclib (HY-101212, Medchemexpress), AI letrozole (CGS 20267; S1235, Selleckchem) and enzalutamide (MDV3100, S1250, Selleckchem) were dissolved in dimethyl-sulfoxide (DMSO, Sigma-Aldrich). CDK4/6i palbociclib (PD332991; PZ0383, Sigma-Aldrich) was dissolved in double-distilled water (ddH₂O).