Sibub1

Depletion of endogenous proteins was achieved through transfection of single small interfering RNA (siRNA) with RNAiMAX (Invitrogen) according to manufacturer’s instructions. Following siRNAs treatments were performed in this study: 50 nM siBUB1 (Dharmacon, 5’-GGUUGCCAACACAAGUUCU3’) for 24 h, 100 nM siBUBR1 (Dharmacon, 5’-CGGGCAUUUGAAUAUGAAA-3’) for 24 h, 60 nM siCENP-E (Dharmacon, 5’-AAGGCUACAAUGGUACUAUAU-3’) for 24 h, 50 nM siCENP-F (Dharmacon, 5’-CAAAGACCGGUGUUACCAAG-3’; 5’-AAGAGAAGACCCCAAGUCAUC-3’) for 24 h, siKNL1 (Invitrogen, HSS183683 5’-CACCCAGUGUCAUACAGCCAAUAUU-3’; HSS125942 5’-UCUACUGUGGUGGAGUUCUUGAUAA-3’; CCCUCUGGAGGAAUGGUCUAAUAAU-3’) for 24 h, siSpindly (Sigma-Aldrich, 5′-GAAAGGGUCUCAAACUGAA-3′) for 48 h, siNdc80C (Sigma-Aldrich, siHec1 5’-GAGUAGAACUAGAAUGUGA-3’; siSpc24 5’-GGACACGACAGUCACAAUC-3’; siSpc25 5’-CUACAAGGAUUCCAUCAAA-3’) for 48 h, 100 nM siZwilch (SMART pool from Dharmacon, L-019377 -00-0005) for 72 h.
Unless indicated, Nocodazole (Sigma-Aldrich) was used at 3.3 μM, RO3306 (Calbiochem) at 9 μM, MG-132 (Calbiochem) at 10 μM, and Reversine (Cayman Chem.) at 500 nM.