KEY RESOURCES TABLE.
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
|
| ||
| Antibodies | ||
|
| ||
| Rabbit anti-ATF4 at 1:250 | Cell Signaling Technology | 11815 |
| Rabbit anti-phospho-S6 (Ser240/244) at 1:250 | Cell Signaling Technology | 2215 |
| Rabbit anti-Cyclin D1 (SP4) at 1:500 | Lab Vision | RM-9104-S0 |
| Rabbit anti-p21 at 1:250 | Cell Signaling Technology | 2947 |
| Mouse anti-p27 at 1:250 | BD Biosciences | 610241 |
| Rabbit anti-phospho-Rb (Ser807/811) (D20B12) at 1:500 | Cell Signaling Technology | 8516 |
| Alexa Fluor 546 anti-mouse secondary at 1:500 | Thermo Fisher | A-11030 |
| Alexa Fluor 647 anti-mouse secondary at 1:500 | Thermo Fisher | A-21236 |
| Alexa Flour 546 anti-rabbit secondary at 1:500 | Thermo Fisher | A-11035 |
| Alexa Fluor 647 anti-rabbit secondary at 1:500 | Thermo Fisher | A-21245 |
|
| ||
| Chemicals, Peptides, and Recombinant Proteins | ||
|
| ||
| Hoechst | Biotium | 40046 |
| Mek inhibitor | Selleckchem | PD-0325091 |
| Cycloheximide | Enzo Life Science | ALX-380-269-G001 |
| Chloroquine | Sigma-Aldrich | AAJ6445914 |
| 5-(N-Ethyl-N-isopropyl)-Amiloride | Selleckchem | S9849 |
| Trimethoprim | Thermo Fisher | AAJ6305303 |
| DharmaFECT 1 | Dharmacon | T-2001-02 |
| DMEM/F-12 | Thermo Fisher | 11039047 |
| DMEM/F-12 w/o Amino Acids, L-Glutamine, Glucose, Sodium Pyruvate | USBiological | D9807-11 |
| Horse Serum | Life Tech | 16050–122 |
| Dialyzed Equine Serum | Valley Biomedical | AS3053 |
| EGF | Peprotech | AF-100-15 |
| Hydrocortisone | Sigma-Aldrich | H0888 |
| Insulin | Sigma-Aldrich | I1882 |
| Cholera Toxin | Sigma-Aldrich | C8052 |
| Collagen | Advanced BioMatrix | 5015 |
|
| ||
| Critical commercial assays | ||
|
| ||
| ViewRNA™ ISH Cell Assay Kit | Thermo Fisher | QVC0001 |
| CCND1 FISH probes | Thermo Fisher | VA6-16943 |
|
| ||
| Experimental models: Cell lines | ||
|
| ||
| MCF10A | ATCC | CRL-10317 |
| RPE-hTERT | ATCC | CRL-4000 |
| MCF10A H2B-mTurq DHB-mVenus DHFR-mCherry | This study | N/A |
| MCF10A H2B-mTurq DHB-mVenus DHFR-mCherry-Cyclin D1 | (Yang et al., 2017)32 | N/A |
| mCitrine-CCND1 MCF10A | (Gookin et al., 2017)25 | N/A |
| mCitrine-CDKN1A MCF10A | (Moser et al., 2018)21 | N/A |
| p21-null MCF10A | (Bachman et al., 2004)36 | N/A |
| Oligonucleotides | ||
| siCDKN1A | IDT | hs.Ri.CDKN1A.13.1 hs.Ri.CDKN1A.13.3 |
| siCDKN1B | IDT | hs.Ri.CDKN1B.13.1 hs.Ri.CDKN1B.13.2 |
| siControl | Horizon Discovery | D-001810-02-05 |
|
| ||
| Other | ||
|
| ||
| 96-well glass bottom plate | Cellvis | Cat#P96-1.5H-N |
|
| ||
| Software and algorithms | ||
|
| ||
| MATLAB R2020a | Mathworks | N/A |
| Adobe Illustrator 2022 | Adobe | N/A |
| ImageJ | Fiji | N/A |
| Tracking code for MCF10A cells | (Cappelletal., 2016)40 | https://github.com/scappell/Cell_tracking |
| Tracking code for RPE hTERT cells | (Tian et al., 2020)41 | https://github.com/tianchengzhe/EllipTrack |
| Microscopy image analysis scripts for plotting tracedata mat files | Spencer Lab | https://doi.org/10.5281/zenodo.10052240 |