|
| Antibodies |
|
| Mouse monoclonal anti-Tubulin (clone DM1A) |
Sigma-Aldrich |
Cat# T9026; RRID: AB_477593 |
| Rabbit polyclonal PRC1 |
Laboratory of Tarun M Kapoor, The Rockefeller University, New York, NY.68
|
N.C |
| Mouse monoclonal 9G3/Hec1 |
Abcam |
Cat# ab3613; RRID: AB_303949 |
| Mouse Alexa Fluor 594 |
Thermo Fisher Scientific |
Cat# A-11032; RRID: AB_2534091 |
| Goat anti-Mouse IgG Alexa Fluor 647 |
Thermo Fisher Scientific |
Cat# A-21236; RRID: AB_2535805 |
| Goat anti-Rabbit Alexa Fluor 594 |
Thermo Fisher Scientific |
Cat# A-11012; RRID: AB_141359 |
| Goat anti-Mouse IgG1 (γ1) Alexa Fluor 488 |
Thermo Fisher Scientific |
Cat# A-21121; RRID: AB_2535764 |
| Goat anti-Mouse IgG2a Alexa Fluor 594 |
Thermo Fisher Scientific |
Cat# A-21135; RRID: AB_2535774 |
|
| Chemicals, peptides, and recombinant proteins |
|
| GSK-923295 |
MedChemExpress |
Cat# HY-10299 |
| PIPES |
Sigma-Aldrich |
E006757; CAS: 5625-37-6 |
| EGTA |
Sigma-Aldrich |
E4378; CAS: 67-42-5 |
| MgCl2
|
Sigma-Aldrich |
CAS 7791-18-6 |
| Triton X-100 |
Sigma-Aldrich |
X-100; CAS No: 9036-19-5 |
| Glutaraldehyde |
Sigma-Aldrich |
G5882; CAS: 111-30-8 |
| Paraformaldehyde |
Electron Microscopy Sciences |
Cat# 15714 |
| EDTA |
Sigma-Aldrich |
E-5134; CAS: 6381-92-6 |
| Phosphate-buffered saline, pH 7.2 |
Thermo Fisher Scientific |
Cat# 20012050 |
| Tween 20 |
Sigma-Aldrich |
P1379; CAS 9005-64-5 |
| Sodium Borohydride |
Sigma-Aldrich |
452882; CAS: 16940-66-2 |
| Hoechst 33342 |
Molecular Probes |
Cat# H3570 |
| CaCl2
|
Acros Organics |
AC123350025; CAS 10035-04-8 |
| HEPES |
Sigma-Aldrich |
H4034; CAS 7365-45-9 |
| KCl |
Fisher |
Cat# P217-3; CAS 7447-40-7 |
| NaCl |
Fisher |
Cat# S640-3; CAS 7647-14-5 |
| Na2HPO4
|
Sigma |
Cat# S374-3; CAS 7558-79-4 |
| Dextrose |
Fisher |
Cat# BP350-1; CAS 50-99-7 |
| Polybrene |
Sigma-Aldrich |
Cat# H9268; CAS: 28728-55-4 |
| Blasticidin |
InvivoGen |
ant-bl-05 |
| Puromycin |
Sigma-Aldrich |
P7255; CAS 58-58-2 |
| Doxycycline Hyclate |
Sigma-Aldrich |
D9891; CAS: 24390-14-5 |
|
| Critical commercial assays |
|
| SiR-Tubulin Kit |
Spirochrome AG |
CY-SC002 |
| Lipofectamine 2000 |
ThermoFisher |
11668027 |
|
| Deposited data |
|
| Kinetochore tracking data |
This study |
https://doi.org/10.5281/zenodo.5803448
|
|
| Experimental models: Cell lines |
|
| Human: hTERT-RPE-1(retinal pigmented epithelium, female) co-expressing CENP-A-GFP and centrin1-GFP |
Laboratory of Alexey Khodjakov, Wadsworth Center, New York State Department of Health, Albany, NY.19
|
N/A |
| Human: hTERT-RPE-1 expressing GFP-PRC1 |
Laboratory of Tarun M Kapoor, The Rockefeller University, New York, NY.68
|
N/A |
| Human: hTERT-RPE-1 expressing Sh-PRC1 RPE1 |
Laboratory of Tarun M Kapoor, The Rockefeller University, New York, NY.29
|
N/A |
| Human: hTERT RPE KNTC1-/-
|
Laboratory of Prasad V. Jallepalli, loan Kettering Institute, Memorial Sloan Kettering Cancer Center, New York, NY.12
|
N/A |
| Human: hTERT-RPE-1 TetON Sh-PRC1 RPE1 |
This study |
N/A |
| Human: Ampho-293 (embryonic kidney, female) |
Clontech |
631505 |
| Human: hTERT-RPE1 RodΔ/Δ co-expressing CENP-A-GFP and centrin1-GFP |
This study |
N/A |
|
| Oligonucleotides |
|
| shRNA targeting sequence: PRC1; 5-GTGATTGAGGCAATTCGAG-3′ |
Laboratory of Tarun M Kapoor, The Rockefeller University, New York, NY.29
|
N/A |
|
| Recombinant DNA |
|
| pMSCVblast vector69
|
Addgene |
Addgene # 7508 |
|
| Software and algorithms |
|
| ImageJ/Fiji |
NIH |
https://imagej.nih.gov/ij/
|
| MATLAB, R2021a |
Mathworks |
https://www.mathworks.com
|
| Imaris |
Oxford Instruments |
https://imaris.oxinst.com
|
| Adobe Suite CC 2021 |
Adobe |
https://www.adobe.com
|
| SoftWoRx 5.0 |
Applied Precision |
http://www.api.com/softworx.asp
|
| Simulation code |
This study |
https://doi.org/10.5281/zenodo.5804405
|
