The RZZ complex integrates spindle checkpoint maintenance with dynamic expansion of unattached kinetochores

Jose-Antonio Rodriguez-Rodriguez; Kara L. McKinley; Vitali Sikirzhytski; Jennifer Corona; John Maciejowski; Alexey Khodjakov; Iain M. Cheeseman; Prasad V. Jallepalli

Cell lines used in this study are described in the Key Resource Table. HeLa (human cervical adenocarcinoma, female) and HEK293 (human embryonic kidney, female) derivatives were grown at 37°C in Dulbecco's modified eagle medium (DMEM) with 10% tetracycline free fetal bovine serum, 100 U/ml penicillin, and 100 U/ml streptomycin. hTERT-RPE (human retinal pigment epithelium, female) derivatives were grown at 37°C in a 1:1 mixture of DMEM and Ham's F-12 medium with 10% fetal bovine serum, 100 U/ml penicillin, and 100 U/ml streptomycin, and 2.5 mM L-glutamine. Integrants were selected using hygromycin (0.2 mg/ml), picked as single colonies, and induced with doxycycline (0.8 μ g/ml). mCherry-Mis12-Mad1 [26] was cloned into a piggyBac vector containing a doxycycline-inducible promoter (tetON) and constitutively expressing reverse tetracycline transactivator (rtTA) and neomycin phosphotransferase All rights reserved. No reuse allowed without permission.