Inducing Tetraploid Cell State

Cells were grown in DMEM medium (RPE-1) or RPMI 1640 medium (HL60, K562, KG1α, and THP1) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin and maintained at 37° C with 5% CO2. For experiments where PLK4 expression was induced, media was supplemented with 2 μg/mL Doxycycline for 36 hours. Alternatively, cells were treated with 30 μM Cytochalasin B for 16 h to induce cytokinesis failure and generate tetraploid cells with double the normal centrosome number [8 (link)]. Drug treatments to subsequently inhibit AurA kinase (alisertib; MLN8054, Aurora A inhibitor 1, MK-5108 (VX-689): Selleckchem) were performed at the indicated concentrations for 16-18 hours for immunofluorescence and FACs analysis, or as otherwise indicated. Unless otherwise noted, all experiments to inhibit Aurora A kinase were performed with the highly specific Aurora A kinase inhibitor alisertib at a concentration of 100 nM.