KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| antibodies | ||
| Mouse monoclonal anti-Flag | Sigma-Aldrich | F3165 |
| Rabbit polyclonal anti-Flag | Sigma-Aldrich | F7425 |
| Mouse monoclonal anti-GFP | Abclonal | AE012 |
| Rabbit polyclonal anti-GFP | ABCAM | ab290 |
| Rabbit polyclonal anti-GFP | Invitrogen | A11122 |
| Rabbit polyclonal anti-GFP | This study | ANTI #76 (Home-Made antibody) |
| Mouse monoclonal anti-CENP-A | Stressgen, Enzo Life Sciences | KAM-CC006 |
| Rabbit polyclonal anti-CENP-A | Upstate | 07–574 |
| Mouse monoclonal anti-CENP-B | Novus Biologicals | H00001059-B01P |
| Rabbit polyclonal anti-CENP-B | abcam | ab25734 |
| Rabbit polyclonal anti-HJURP | Proteintech Group Inc. | 15283–1-AP |
| Rabbit polyclonal anti-GAPDH | ABCAM | ab37168 |
| Rabbit polyclonal anti-GAPDH | Invitrogen | PA1987 |
| Rabbit polyclonal anti-Ubiquitin | Bethyl Laboratories | A300–317A-1 |
| Affinity-purified secondary antibody Alexa Fluor 488 Goat anti-Mouse IgG | Invitrogen | A11001 |
| Affinity-purified secondary antibody Alexa Fluor 594 Goat anti-Mouse IgG | Invitrogen | A11005 |
| Affinity-purified secondary antibody Alexa Fluor 488 Goat anti-Rabbit IgG | Invitrogen | A11008 |
| Affinity-purified secondary antibody Alexa Fluor 594 Goat anti-Rabbit IgG | Invitrogen | A11012 |
| Affinity-purified secondary antibody Alexa Fluor Plus 488 Goat anti-Mouse IgG | Invitrogen | A32723 |
| Affinity-purified secondary antibody Alexa Fluor Plus 555 Goat anti-Rabbit IgG | Invitrogen | A32732 |
| Chemicals, Peptides, and Recombinant Proteins | ||
| Fetal bovine serum (FBS) | Gibco, Invitrogen | 10082–147 |
| 100 units penicillin/100 μg streptomycin | Gibco, Invitrogen | 15140–148 |
| High-glucose Dulbecco’s modified Eagle’s (DMEM) medium | BioWhittaker | 12–604f |
| Lipofectamine 3000 | Invitrogen | L3000–008 |
| Polyethylenimine (PEI) | Polysciences, Inc. | 23966–2 |
| Complete EDTA-free protease inhibitor cocktail | Roche | 11-873-580-001 |
| Restore Western Blot Stripping Buffer | Thermo Scientific | PI21059 |
| Protein A Sepharose CL-4B | GE Healthcare | 17-0963-03 |
| Anti-Flag M2 affinity gel | Sigma-Aldrich | A2220–1 ML |
| DAPI | Sigma-Aldrich | D9542 |
| Crystal violet solution | SigmaI-Aldrich | HT90132–1L |
| DMEM:DMEM/F12 medium | ATCC | 30–2006 |
| Experimental Models: Cell Lines (sex of cell is indicated.) | ||
| CENP-A−/f RPE-1 cells (female) | (Fachinetti et al., 2017), This study | Dr. Daniele Fachinetti, Dr. Don W. Cleveland |
| CENP-AF/+ RPE-1 cells (female) | (Fachinetti et al., 2017), This study | Dr. Daniele Fachinetti, Dr. Don W. Cleveland |
| HeLa cells (female) | (Niikura and Kitagawa, 2016; Niikura et al., 2015; Niikura et al., 2017) | N/A |
| 293T cells (female) | (Niikura et al., 2016) | N/A |
| Recombinant DNA | ||
| pEGFP-C1 | This study | B161 |
| pEGFP-C1-human CENP-A WT | This study | Dr. Yamini Dalal, B3128 |
| pEGFP-C1-human CENP-A K124A | This study | Dr. Yamini Dalal, B3129 |
| pEGFP-C1-human CENP-A K124Q | This study | Dr. Yamini Dalal, B3130 |
| pcDNA3 | This study | B1512 |
| pcDNA3-Flag-human CENP-A WT | This study | B2854 |
| pcDNA3-Flag-human CENP-A K124R | This study | B2829 |
| pcDNA3-HA-human CENP-A WT | This study | B2852 |
| pcDNA3-HA-human CENP-A K124R | This study | B2836 |
| pBabe-EYFP | This study | B3182 |
| pBabe-EYFP-human CENP-A WT | (Fachinetti et al., 2017), This study | Dr. Daniele Fachinetti, Dr. Don W. Cleveland, B3161 |
| pBabe-EYFP-human CENP-A K124R | (Fachinetti et al., 2017), This study | Dr. Daniele Fachinetti, Dr. Don W. Cleveland, B3164 |
| pQCXIP-EYFP | This study | B3252 |
| pQCXIP-EYFP-human CENP-A WT | This study | B3254 |
| pQCXIP-EYFP-human CENP-A K124R | This study | B3256 |
| pBabe-Flag-human CENP-A WT | This study | B3237 |
| pBabe-Flag-human CENP-A K124R | This study | B3236 |
| pBabe-untagged human CENP-A WT | This study | B3234 |
| pBabe-untagged human CENP-A K124R | This study | B3235 |
| pBabe-Flag-human CENP-A K124R-UbK48R | This study | B3271 |
| pBabe-untagged human CENP-A K124R-UbK48R | This study | B3270 |
| pCGN-HA-Ubiquitin | (Niikura and Kitagawa, 2016; Niikura et al., 2015; Niikura et al., 2017), This study | B2806 |
| pVSV-G | (Fachinetti et al., 2017), This study | Dr. Daniele Fachinetti, Dr. Don W. Cleveland, B3163 |
| pCMV-VSV-G | This study | B1197 |
| pGP | This study | Dr. Kenji Tago (Jichi Medical Univeristy, Japan), B3189 |
| pCI-VSVG | This study | Dr. Kenji Tago (Jichi Medical Univeristy, Japan), B3190 |
| pPAM | This study | B3001 |
| pBabe-puro | (Niikura et al., 2016), This study | Dr. Amruta Ashtekar, Dr. Lawrence S. Kirschner, B3028 |
| pBabe-puro-Cre | (Niikura et al., 2016), This study | Dr. Amruta Ashtekar, Dr. Lawrence S. Kirschner, B3027 |
| psPAX2 | (Niikura et al., 2016), This study | Dr. John Thompson, Dr. Gustavo W. Leone, B3031 |
| pMD2.g | (Niikura et al., 2016), This study | Dr. John Thompson, Dr. Gustavo W. Leone, B3032 |
| Software and Algorithms | ||
| Odyssey CLx Infrared Imaging System | LI-COR Biosciences | https://www.licor.com/bio/products/imaging_systems/ |
| Odyssey Fc Imaging System | LI-COR Biosciences | https://www.licor.com/bio/products/imaging_systems/ |
| FluorChem M system | Proteinsimple | https://www.proteinsimple.com/fluorchem_m.html |
| Image Studio Software | LI-COR Biosciences | https://www.licor.com/bio/products/imaging_systems/ |
| Quantity One 1-D Analysis Software | Bio-Rad | http://www.bio-rad.com/zh-cn/product/quantity-one-1-d-analysis-software?ID=1de9eb3a-1eb5-4edb-82d2-68b91bf360fb |
| ImageJ software | NIH | https://imagej.nih.gov/ij/ |
| Olympus FLUOVIEW FV3000 confocal laser scanning microscope | Olympus | https://www.olympus-global.com/news/2016/nr160405fv3000e.html |
| Acquisition FV31S-SW software | Olympus | https://www.olympus-lifescience.com.cn/en/support/downloads/#!dlOpen=%23detail847250519 |
| Analysis FV31S-DT software | Olympus | https://www.olympus-lifescience.com.cn/en/support/downloads/#!dlOpen=%23detail847250519 |
| cellSens Dimension software Ver. 1. 18 | Olympus | https://www.olympus-lifescience.com.cn/en/software/cellsens/ |
| DM IRE2 motorized fluorescence microscope | Leica | https://www.leica-microsystems.com/applications/life-science/fluorescence/ |
| ARC LAMP power supply HBO100 DC IGN | Ludl Electronic Products, Ltd. | N/A |
| ORCA-ER high-resolution digital CCD camera | Hamamatsu | N/A |
| Openlab version 5.5.2 Scientific Imaging Software | Improvision, PerkinElmer | http://www.perkinelmer.com/lab-products-and-services/resources/cellular-imaging-software-downloads.html |
| Velocity version 6.3 3D Image Analysis Software | Improvision, PerkinElmer | http://www.perkinelmer.com/lab-products-and-services/resources/cellular-imaging-software-downloads.html |
| Molecular Imager Versadoc MP4000 System | Bio-Rad | http://www.bio-rad.com/zh-cn/sku/1708640-molecular-imager-versadoc-mp-4000-system?ID=1708640 |
| Quantity One 1-D Analysis Software | Bio-Rad | http://www.bio-rad.com/zh-cn/product/quantity-one-1-d-analysis-software?ID=1de9eb3a-1eb5-4edb-82d2-68b91bf360fb |
| OpenCFU | https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0054072 | http://opencfu.sourceforge.net/ |
| Other | ||
| Immobilon-FL PVDF Transfer Membrane | EMD Millipore | IPVH00010 |
| NuPAGE™ 4–12% Bis-Tris Protein Gels | Thermo Fisher | NP0335BOX |
| Coomassie brilliant blue R-250 | BBI Life Sciences | CAS 6104-59-2 |