Key resources table.
| Reagent type (species) or resource |
Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Cell line (Homo sapiens) | MCF10A | ATCC | CRL-10317, RRID:CVCL_0598 | |
| Cell line (Homo sapiens) | MCF10A + PB-NES ZapCV2 and PB-H2B- mCherry (stable) | This paper | ||
| Cell line (Homo sapiens) | MCF10A + DHB-Venus and H2B-mTurqoise (stable) | Laboratory of Sabrina Spencer | PMCID:PMC4001917 | |
| Cell line (Homo sapiens) | MCF10A p21-/- + DHB-Venus and H2B-mTurqoise (stable) | Laboratory of Sabrina Spencer | PMCID:PMC4001917 | |
| Recombinant DNA reagent | PB-NES-ZapCV2 (plasmid used for stable cell line generation) | Published in previous Palmer lab paper PMID: 27959493 | Cytosolic FRET sensor in Piggybac vector | |
| Recombinant DNA reagent | PB-H2B-mCherry (plasmid used for stable cell line generation) | This paper | H2B-mCherry subcloned from H2B-mCherry CSII-EF (PMCID:PMC4001917) into PB510B1 Piggybac vector (System Biosciences) with primers listed below | |
| Sequence-based reagent | Forward primer to subclone H2B-mCherry into PB plasmid (PB510B1, System Biosciences) | IDT | 5’-GGAATT GAATTCGCT ACCGGTCTC GAGAC-3’ (EcoRI site for cloning) | |
| Sequence-based reagent | Reverse primer to subclone H2B-mCherry into PB plasmid (PB510B1, System Biosciences) | IDT | 5’-GATTAT GATCTAGAG TCGCGG CCGCCT G-3’ (NotI site for cloning) | |
| Antibody | Anti-Phospho-Rb Ser 807/811 (rabbit monoclonal) | Cell Signalling Technology | D20B12 XP, Lot8, RRID:AB_11178658 | 1:500 (FACS) 1:100 (IF) |
| Antibody | Anti-p21 Waf1/Cip1 (rabbit monoclonal) | Cell Signalling Technology | 2947S, Lot 10, RRID:AB_823586 | 1:200 (IF) |
| Antibody | Anti-53BP1 (mouse monoclonal) | BD Biosciences | 612523, Lot 6217571, RRID:AB_399824 | 1:200 (IF) |
| Antibody | Anti-RPA2 (mouse monoclonal) | Abcam | Abcam, ab2175, Lot GR3224197-5), RRID:AB_302873 | 1:250 (IF) |
| Antibody | AlexaFluor 488 (donkey anti rabbit) | Abcam | ab150073, Lot GR328726-1, RRID:AB_2636877 | 1:500 for pRb (FACS) |
| Antibody | AlexaFluor 488 (goat anti mouse) | Thermo Fisher Scientific | A11029, Lot 1789729,RRID:AB_138404 | 1:500 for 53BP1 and RPA2 (IF) |
| Antibody | AlexaFluor 647 (goat anti rabbit) |
Thermo Fisher Scientific | A21244, Lot 1156625, RRID:AB_141663 | 1:500 for pRb (IF) |
| Antibody | AlexaFluor 568 (donkey anti rabbit) | Thermo Fisher Scientific | A10042, Lot 1134929, RRID:AB_2534017 | 1:500 for p21 in (IF) |
| Commercial assay or kit | Click-IT EdU AlexaFluor 647 HCS Assay | Thermo Fisher Scientific | C10356 | |
| Commercial assay or kit | FxyCycle PI/RNase Staining Solution | Thermo Fisher Scientific | F10797 | |
| Chemical compound, drug | TRIS(2-PYRIDYLMETHYL)AMINE 98% (TPA) | Sigma-Aldrich | 723134 | |
| Chemical compound, drug | N,N,N′,N′-Tetrakis(2-pyridylmethyl)ethylenediamine (TPEN) | Sigma-Aldrich | P4413 | |
| Chemical compound, drug | Zinc chloride, anhydrous, 99.95% (metals basis) | Alfa Aesar | 87900 | |
| Chemical compound, drug | Chelex-100, sodium form | Sigma-Aldrch | C7901 | |
| Chemical compound, drug | Neocuproine hydrochloride Monohydrate | Fluka | 72090 | |
| CF4 | Copper-Fluor-4 | Christopher Chang Lab | PMCID: PMC6008210 | |
| Software | BD FACSDiva | BD Biosciences | Version 8 | |
| Software | MATLAB | Mathworks | V R2017b VR2018a |
|
| Software | KaleidaGraph | Synergy | Version 4.02 | |
| Software | Prism | GraphPad Software | Version 8.2.0 | |
| Other (Code) | Cell segmentation and tracking pipeline generated in Matlab | Desposited: https://biof-git.colorado.edu/biofrontiers-imaging/palmer-zinc-cell-cycle | ||
| Other | Hoechst 33258 | Sigma-Aldrich | 861405 | Used at 0.1 µg/mL in PBS |