KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Mouse monoclonal anti-PCNA (SR) | Santa Cruz | Cat# sc-56, RRID: AB_628110 |
| Rabbit monoclonal anti-MCM6 (AlexaFluor 568 conjugated, SR) | Abcam | Cat# ab211916, RRID: N/A |
| Rabbit monoclonal anti-RPA70 (AlexaFluor 647 conjugated, SR) | Abcam | Cat# ab199240, RRID: AB_2687566 |
| Rabbit polyclonal anti-RPA32(phosphor S4+S8) (SR) | Abcam | Cat# ab87277, RRID: AB_1952482 |
| Goat polyclonal anti-Mouse (AlexaFluor 488 conjugated, SR) | Thermal Fisher | Cat# A-11001, RRID: AB_2534069 |
| Goat polyclonal anti-Rabbit (AlexaFluor 647 conjugated, SR) | Thermal Fisher | Cat# A27040, RRID: AB_2536101 |
| Mouse monoclonal anti-γH2AX (IF) | Millipore | Cat# 16-193, RRID: AB_310795 |
| Goat polyclonal anti-Mouse (AlexaFluor 568 conjugated, IF) | Thermal Fisher | Cat# A-11004, RRID: AB_2534072 |
| Mouse monoclonal anti-RPA32 (WB) | Millipore | Cat# MABE285, RRID: AB_11205561 |
| Rabbit polyclonal anti-Myc (WB) | Bethyl | Cat# A190-105A, RRID: AB_67390 |
| Mouse monoclonal anti-β-Tublin (WB) | Sigma-Aldrich | Cat# T8328, RRID: AB_1844090 |
| Mouse polyclonal anti-HRP (WB) | Abcam | Cat# ab181708, RRID: N/A |
| Rabbit polyclonal anti-HRP (WB) | Abcam | Cat# ab34885, RRID: AB_732995 |
| Bacterial and Virus Strains | ||
| Biological Samples | ||
| Chemicals, Peptides, and Recombinant Proteins | ||
| ATR inhibitor, VE-821 | Selleckchem | Cat# S8007, CAS No. 1232410-49-9 |
| CDC7 inhibitor, PHA767491 | Selleckchem | Cat# S2742, CAS No. 942425-68-5 |
| Chk1 inhibitor, UCN-01 | Sigma-Aldrich | Cat# U6508, CAS No. 112953-11-4 |
| Chk2 inhibitor, PV1019 | Sigma-Aldrich | Cat# 220488, CAS No. 516480-79-8 |
| Hydroxyurea | Sigma-Aldrich | Cat# H8627, CAS No. 127-07-1 |
| Hoechst-33342 | Thermo Fisher | Cat# H3570, CAS No. 23491-52-3 |
| DMEM cell culture medium | Thermo Fisher | Cat# 11965 |
| FBS | Gemini Bio | Cat# 100-106 |
| Penicillin-Streptomycin | Thermo Fisher | Cat# 15140 |
| Critical Commercial Assays | ||
| Click reaction kit for EdU and Alexa Fluor 647 azide | Thermo Fisher | Cat# 10640 |
| Lipofectamine RNAiMax reagent | Thermo Fisher | Cat# 13778075 |
| Lipofectamine 3000 transfection kit | Thermo Fisher | Cat# L3000015 |
| Deposited Data | ||
| Representative images, uncropped gels, all the data | This study | DOI: 10.17632/s4b8ztvnxd.1 |
| Experimental Models: Cell Lines | ||
| U2OS | ATCC | Cat# HTB-96, RRID: CVCL_0042 |
| Experimental Models: Organisms/Strains | ||
| Oligonucleotides | ||
| ATRIP FlexiTube GeneSolution | QIAGEN | SI03072496 |
| ON-TARGET plus Human RPA2 siRNA | Dharmacon | SO-2916681G |
| 5′-TGGCGACGGCAGCGAGGCTTTTTTTTTTTTTTTTTTTT-iCy3-TTTTTTTTTTTT-3′-Cy5 (Figure 5) | IDT | N/A |
| 5′-GCCTCGCTGCCGTCGCCA-3′-biotin (Figure 5) | IDT | N/A |
| 5′-TGGCGACGGCAGCGAGGCTTTTTTTTTTTTTTTT-iCy3-TTTTTTTTTTTTTT-3′ (Figure S6) | IDT | N/A |
| Cy5-5′-GCCTCGCTGCCGTCGCCA-3′-biotin (Figure S6) | IDT | N/A |
| Recombinant DNA | ||
| Plasmid pEF6-RPA2-WT-Myc | This study | N/A |
| Plasmid pEF6-RPA2-S33D-Myc | This study | N/A |
| Plasmid pEF6-RPA2-S33A-Myc | This study | N/A |
| Software and Algorithms | ||
| Reconstruction of SMLM (CUDA + Matlab) | CUDA, MathWorks | MATLAB, RRID: SCR_001622 |
| TCF (Matlab) | Yin et al., 2019 | DOI: 10.17632/s4b8ztvnxd.1 |
| FACSCalibur | BD Biosciences | RRID: SCR_000401 |
| ImageJ | Schneider et al., 2012 | https://imagej.nih.gov/ij/ |
| Originlab | OriginLab | https://www.originlab.com/ |
| Other | ||