Mk 5108

Drugs, suppliers, and concentrations used were Barasertib (Aurora B inhibitor; alternative name AZD1152‐HQPS; SelleckChem S1147; 1.11 nM); CHR‐6494 (Haspin inhibitor; MedChem Express HY‐15217; 500 nM); CW069 (HSET inhibitor; SelleckChem S7336; 25.0 μM); Etoposide (Topoisomerase II inhibitor; SelleckChem S1225; 333 nM); GSK461364 (PLK1 inhibitor; SelleckChem S2193; 2.20 nM); GSK923295 (CENP‐E inhibitor; SelleckChem S7090; 3.20 nM); Ispinesib (KIF11 inhibitor; alternative name SB‐715992; SelleckChem S1452; 1.70 nM); MK‐5108 (Aurora A inhibitor; alternative name VX‐689; SelleckChem S2770; 0.576 nM); MK‐8776 (CHK1 inhibitor; alternative name SCH 900776; SelleckChem S2735; 9.00 nM); Paclitaxel (microtubule inhibitor; SelleckChem S1150; 2.67 nM); Vinblastine (microtubule inhibitor; Sigma V1377; 2.40 nM); and YM155 (BIRC5 inhibitor; SelleckChem S1130; 0.540 nM).

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.

Parasite was maintained by in vivo propagation of the parasite material in mice (supplied by Xiamen University Laboratory Animals Center, XMULAC). Mature and developing protoscoleces were collected from parasite material, manually picked under the microscope, and then immediately used for RNA isolation or EdU labeling. In vitro cultivation of metacestode vesicles was performed using host cell conditioned medium as previously described [26 (link)]. The growth of metacestode vesicles and the process of vesicle formation from protoscoleces were examined after 21 days and 14 days of culture, respectively as described by Cheng et al. [27 (link)].
Aurora inhibitors (MLN8237, MLN8054, MK-5108 and AZD1152-HQPA), nocodazole and hydroxyurea were supplied by Selleck Chemicals. Drugs were added into the culture medium at a final concentration as indicated. All of the drug experiments were carried out under axenic culture conditions as described before [26 (link)]. For longer periods of treatment, experiments were performed with exchange of the medium containing the same ingredients every three days.