other:Article Title: PLK1 Phosphorylates WRN at Replication Forks
Article Snippet: Prostate cancer, particularly castration-resistant prostate cancer, remains a serious public health issue.. Androgen signaling inhibitors have emerged as a major treatment approach but with limited success.. Thus, identification of novel treatment targets is of high clinical relevance.
Recombinant:Article Title: CtIP-Mediated Fork Protection Synergizes with BRCA1 to Suppress Genomic Instability upon DNA Replication Stress.
Article Snippet: Capan-1 ATCC Cat#HTB-79; RRID: CVCL_0237 H1299 DOX-inducible BRCA2 shRNA Madalena Tarsounas lab, Zimmer et al., 2016 N/A U2OS Flp-In T-REx Daniel Durocher lab, Al-Hakim et al., 2012 N/A PD20F (FANCD2-deficient) Fanconi Anemia Research Foundation (FARF), Murina et al., 2014 N/A PD20F complemented with FANCD2 WT Fanconi Anemia Research Foundation (FARF), Murina et al., 2014 N/A (Continued on next page) Molecular Cell 72, 1–15.e1–e6, November 1, 2018 e2 .. REAGENT or RESOURCE SOURCE IDENTIFIER Oligonucleotides Table S1 N/A N/A Recombinant DNA pEGFP-C1-CtIP (WT, T847A, S327A) Murina et al., 2014 N/A pEGFP-C1-CtIP N181A This paper N/A pcDNA5/FRT/TO-GFP-CtIP WT Ferretti et al., 2016 N/A pcDNA5/FRT/TO-GFP-CtIP (N181A, N289A/H290A) This paper N/A DR-GFP vector (pHPRT-DRGFP) Addgene #26476 N/A I-SceI expression plasmid (pCBASce) Addgene #26477 N/A DU46129, U6_gRNA MRC PPU Reagents and Services https://mrcppureagents.dundee.ac.uk Software and Algorithms GraphPad Prism7 for Mac OS X GraphPad Software http://www.graphpad.com/scientific- software/prism; RRID: SCR_002798 Fiji Department of Health and Human Services, NIDA https://fiji.sc/#; RRID: SCR_002285 Cell Profiler 2.2.0 Broad Institute http://cellprofiler.org; RRID: SCR_007358 FlowJo X FlowJo Software https://www.flowjo.com/; RRID: SCR_008520 FusionCapt Advance Solo 7 17.02 Vilber Lourmat http://www.vilber.de/ Olympus ScanR Image Analysis Software Version 2.5.1 OLYMPUS Image Analysis Software https://www.olympus-ims.com/pl/ microscope/software/ Spotfire TIBCO Software https://spotfire.tibco.com/; RRID: SCR_008858 Other BD FACSAria III BD Biosciences N/A Fusion Solo S Vilber N/A Leica DMI6000 Leica N/A BD LSR II Fortessa BD Biosciences N/A Attune NxT Flow Cytometer Thermo Fisher Scientific N/A CEF-DR II Variable Angle System Biorad N/A SpectraMax i3 Microplate Reader Molecular Devices N/A Olympus IX83 Olympus N/A Olympus ScanR Screening System Olympus N/A Faxitron Cabinet X-ray System Model RX-650 Faxitron N/A ..
Expressing:Article Title: CtIP-Mediated Fork Protection Synergizes with BRCA1 to Suppress Genomic Instability upon DNA Replication Stress.
Article Snippet: Capan-1 ATCC Cat#HTB-79; RRID: CVCL_0237 H1299 DOX-inducible BRCA2 shRNA Madalena Tarsounas lab, Zimmer et al., 2016 N/A U2OS Flp-In T-REx Daniel Durocher lab, Al-Hakim et al., 2012 N/A PD20F (FANCD2-deficient) Fanconi Anemia Research Foundation (FARF), Murina et al., 2014 N/A PD20F complemented with FANCD2 WT Fanconi Anemia Research Foundation (FARF), Murina et al., 2014 N/A (Continued on next page) Molecular Cell 72, 1–15.e1–e6, November 1, 2018 e2 .. REAGENT or RESOURCE SOURCE IDENTIFIER Oligonucleotides Table S1 N/A N/A Recombinant DNA pEGFP-C1-CtIP (WT, T847A, S327A) Murina et al., 2014 N/A pEGFP-C1-CtIP N181A This paper N/A pcDNA5/FRT/TO-GFP-CtIP WT Ferretti et al., 2016 N/A pcDNA5/FRT/TO-GFP-CtIP (N181A, N289A/H290A) This paper N/A DR-GFP vector (pHPRT-DRGFP) Addgene #26476 N/A I-SceI expression plasmid (pCBASce) Addgene #26477 N/A DU46129, U6_gRNA MRC PPU Reagents and Services https://mrcppureagents.dundee.ac.uk Software and Algorithms GraphPad Prism7 for Mac OS X GraphPad Software http://www.graphpad.com/scientific- software/prism; RRID: SCR_002798 Fiji Department of Health and Human Services, NIDA https://fiji.sc/#; RRID: SCR_002285 Cell Profiler 2.2.0 Broad Institute http://cellprofiler.org; RRID: SCR_007358 FlowJo X FlowJo Software https://www.flowjo.com/; RRID: SCR_008520 FusionCapt Advance Solo 7 17.02 Vilber Lourmat http://www.vilber.de/ Olympus ScanR Image Analysis Software Version 2.5.1 OLYMPUS Image Analysis Software https://www.olympus-ims.com/pl/ microscope/software/ Spotfire TIBCO Software https://spotfire.tibco.com/; RRID: SCR_008858 Other BD FACSAria III BD Biosciences N/A Fusion Solo S Vilber N/A Leica DMI6000 Leica N/A BD LSR II Fortessa BD Biosciences N/A Attune NxT Flow Cytometer Thermo Fisher Scientific N/A CEF-DR II Variable Angle System Biorad N/A SpectraMax i3 Microplate Reader Molecular Devices N/A Olympus IX83 Olympus N/A Olympus ScanR Screening System Olympus N/A Faxitron Cabinet X-ray System Model RX-650 Faxitron N/A ..
Article Title: PALB2 deficiency may sensitize H3K27M-mutant pediatric HGG cells to BMN673/talazoparib
Article Snippet: .. The indicated EJ5 reporter cell lines were transfected with two plasmids, namely, the I-SceI expression vector pCBASce (Addgene #26477) and the DsRed expression vector (Addgene #54493), 48 h later, the percentage of GFP-positive cells among DsRed-positive cells was evaluated by imaging with a Carl Zeiss LSM 710 confocal fluorescence microscope and quantified via FACS on a BD AccuriTM C6 flow cytometer (BD Biosciences, USA) using BD AccuriTM software. .. Subcellular protein fractions were collected with a subcellular fractionation kit (Thermo Fisher Scientific, USA).
Plasmid Preparation:Article Title: CtIP-Mediated Fork Protection Synergizes with BRCA1 to Suppress Genomic Instability upon DNA Replication Stress.
Article Snippet: Capan-1 ATCC Cat#HTB-79; RRID: CVCL_0237 H1299 DOX-inducible BRCA2 shRNA Madalena Tarsounas lab, Zimmer et al., 2016 N/A U2OS Flp-In T-REx Daniel Durocher lab, Al-Hakim et al., 2012 N/A PD20F (FANCD2-deficient) Fanconi Anemia Research Foundation (FARF), Murina et al., 2014 N/A PD20F complemented with FANCD2 WT Fanconi Anemia Research Foundation (FARF), Murina et al., 2014 N/A (Continued on next page) Molecular Cell 72, 1–15.e1–e6, November 1, 2018 e2 .. REAGENT or RESOURCE SOURCE IDENTIFIER Oligonucleotides Table S1 N/A N/A Recombinant DNA pEGFP-C1-CtIP (WT, T847A, S327A) Murina et al., 2014 N/A pEGFP-C1-CtIP N181A This paper N/A pcDNA5/FRT/TO-GFP-CtIP WT Ferretti et al., 2016 N/A pcDNA5/FRT/TO-GFP-CtIP (N181A, N289A/H290A) This paper N/A DR-GFP vector (pHPRT-DRGFP) Addgene #26476 N/A I-SceI expression plasmid (pCBASce) Addgene #26477 N/A DU46129, U6_gRNA MRC PPU Reagents and Services https://mrcppureagents.dundee.ac.uk Software and Algorithms GraphPad Prism7 for Mac OS X GraphPad Software http://www.graphpad.com/scientific- software/prism; RRID: SCR_002798 Fiji Department of Health and Human Services, NIDA https://fiji.sc/#; RRID: SCR_002285 Cell Profiler 2.2.0 Broad Institute http://cellprofiler.org; RRID: SCR_007358 FlowJo X FlowJo Software https://www.flowjo.com/; RRID: SCR_008520 FusionCapt Advance Solo 7 17.02 Vilber Lourmat http://www.vilber.de/ Olympus ScanR Image Analysis Software Version 2.5.1 OLYMPUS Image Analysis Software https://www.olympus-ims.com/pl/ microscope/software/ Spotfire TIBCO Software https://spotfire.tibco.com/; RRID: SCR_008858 Other BD FACSAria III BD Biosciences N/A Fusion Solo S Vilber N/A Leica DMI6000 Leica N/A BD LSR II Fortessa BD Biosciences N/A Attune NxT Flow Cytometer Thermo Fisher Scientific N/A CEF-DR II Variable Angle System Biorad N/A SpectraMax i3 Microplate Reader Molecular Devices N/A Olympus IX83 Olympus N/A Olympus ScanR Screening System Olympus N/A Faxitron Cabinet X-ray System Model RX-650 Faxitron N/A ..
Software:Article Title: CtIP-Mediated Fork Protection Synergizes with BRCA1 to Suppress Genomic Instability upon DNA Replication Stress.
Article Snippet: Capan-1 ATCC Cat#HTB-79; RRID: CVCL_0237 H1299 DOX-inducible BRCA2 shRNA Madalena Tarsounas lab, Zimmer et al., 2016 N/A U2OS Flp-In T-REx Daniel Durocher lab, Al-Hakim et al., 2012 N/A PD20F (FANCD2-deficient) Fanconi Anemia Research Foundation (FARF), Murina et al., 2014 N/A PD20F complemented with FANCD2 WT Fanconi Anemia Research Foundation (FARF), Murina et al., 2014 N/A (Continued on next page) Molecular Cell 72, 1–15.e1–e6, November 1, 2018 e2 .. REAGENT or RESOURCE SOURCE IDENTIFIER Oligonucleotides Table S1 N/A N/A Recombinant DNA pEGFP-C1-CtIP (WT, T847A, S327A) Murina et al., 2014 N/A pEGFP-C1-CtIP N181A This paper N/A pcDNA5/FRT/TO-GFP-CtIP WT Ferretti et al., 2016 N/A pcDNA5/FRT/TO-GFP-CtIP (N181A, N289A/H290A) This paper N/A DR-GFP vector (pHPRT-DRGFP) Addgene #26476 N/A I-SceI expression plasmid (pCBASce) Addgene #26477 N/A DU46129, U6_gRNA MRC PPU Reagents and Services https://mrcppureagents.dundee.ac.uk Software and Algorithms GraphPad Prism7 for Mac OS X GraphPad Software http://www.graphpad.com/scientific- software/prism; RRID: SCR_002798 Fiji Department of Health and Human Services, NIDA https://fiji.sc/#; RRID: SCR_002285 Cell Profiler 2.2.0 Broad Institute http://cellprofiler.org; RRID: SCR_007358 FlowJo X FlowJo Software https://www.flowjo.com/; RRID: SCR_008520 FusionCapt Advance Solo 7 17.02 Vilber Lourmat http://www.vilber.de/ Olympus ScanR Image Analysis Software Version 2.5.1 OLYMPUS Image Analysis Software https://www.olympus-ims.com/pl/ microscope/software/ Spotfire TIBCO Software https://spotfire.tibco.com/; RRID: SCR_008858 Other BD FACSAria III BD Biosciences N/A Fusion Solo S Vilber N/A Leica DMI6000 Leica N/A BD LSR II Fortessa BD Biosciences N/A Attune NxT Flow Cytometer Thermo Fisher Scientific N/A CEF-DR II Variable Angle System Biorad N/A SpectraMax i3 Microplate Reader Molecular Devices N/A Olympus IX83 Olympus N/A Olympus ScanR Screening System Olympus N/A Faxitron Cabinet X-ray System Model RX-650 Faxitron N/A ..
Article Title: PALB2 deficiency may sensitize H3K27M-mutant pediatric HGG cells to BMN673/talazoparib
Article Snippet: .. The indicated EJ5 reporter cell lines were transfected with two plasmids, namely, the I-SceI expression vector pCBASce (Addgene #26477) and the DsRed expression vector (Addgene #54493), 48 h later, the percentage of GFP-positive cells among DsRed-positive cells was evaluated by imaging with a Carl Zeiss LSM 710 confocal fluorescence microscope and quantified via FACS on a BD AccuriTM C6 flow cytometer (BD Biosciences, USA) using BD AccuriTM software. .. Subcellular protein fractions were collected with a subcellular fractionation kit (Thermo Fisher Scientific, USA).
Microscopy:Article Title: CtIP-Mediated Fork Protection Synergizes with BRCA1 to Suppress Genomic Instability upon DNA Replication Stress.
Article Snippet: Capan-1 ATCC Cat#HTB-79; RRID: CVCL_0237 H1299 DOX-inducible BRCA2 shRNA Madalena Tarsounas lab, Zimmer et al., 2016 N/A U2OS Flp-In T-REx Daniel Durocher lab, Al-Hakim et al., 2012 N/A PD20F (FANCD2-deficient) Fanconi Anemia Research Foundation (FARF), Murina et al., 2014 N/A PD20F complemented with FANCD2 WT Fanconi Anemia Research Foundation (FARF), Murina et al., 2014 N/A (Continued on next page) Molecular Cell 72, 1–15.e1–e6, November 1, 2018 e2 .. REAGENT or RESOURCE SOURCE IDENTIFIER Oligonucleotides Table S1 N/A N/A Recombinant DNA pEGFP-C1-CtIP (WT, T847A, S327A) Murina et al., 2014 N/A pEGFP-C1-CtIP N181A This paper N/A pcDNA5/FRT/TO-GFP-CtIP WT Ferretti et al., 2016 N/A pcDNA5/FRT/TO-GFP-CtIP (N181A, N289A/H290A) This paper N/A DR-GFP vector (pHPRT-DRGFP) Addgene #26476 N/A I-SceI expression plasmid (pCBASce) Addgene #26477 N/A DU46129, U6_gRNA MRC PPU Reagents and Services https://mrcppureagents.dundee.ac.uk Software and Algorithms GraphPad Prism7 for Mac OS X GraphPad Software http://www.graphpad.com/scientific- software/prism; RRID: SCR_002798 Fiji Department of Health and Human Services, NIDA https://fiji.sc/#; RRID: SCR_002285 Cell Profiler 2.2.0 Broad Institute http://cellprofiler.org; RRID: SCR_007358 FlowJo X FlowJo Software https://www.flowjo.com/; RRID: SCR_008520 FusionCapt Advance Solo 7 17.02 Vilber Lourmat http://www.vilber.de/ Olympus ScanR Image Analysis Software Version 2.5.1 OLYMPUS Image Analysis Software https://www.olympus-ims.com/pl/ microscope/software/ Spotfire TIBCO Software https://spotfire.tibco.com/; RRID: SCR_008858 Other BD FACSAria III BD Biosciences N/A Fusion Solo S Vilber N/A Leica DMI6000 Leica N/A BD LSR II Fortessa BD Biosciences N/A Attune NxT Flow Cytometer Thermo Fisher Scientific N/A CEF-DR II Variable Angle System Biorad N/A SpectraMax i3 Microplate Reader Molecular Devices N/A Olympus IX83 Olympus N/A Olympus ScanR Screening System Olympus N/A Faxitron Cabinet X-ray System Model RX-650 Faxitron N/A ..
Article Title: PALB2 deficiency may sensitize H3K27M-mutant pediatric HGG cells to BMN673/talazoparib
Article Snippet: .. The indicated EJ5 reporter cell lines were transfected with two plasmids, namely, the I-SceI expression vector pCBASce (Addgene #26477) and the DsRed expression vector (Addgene #54493), 48 h later, the percentage of GFP-positive cells among DsRed-positive cells was evaluated by imaging with a Carl Zeiss LSM 710 confocal fluorescence microscope and quantified via FACS on a BD AccuriTM C6 flow cytometer (BD Biosciences, USA) using BD AccuriTM software. .. Subcellular protein fractions were collected with a subcellular fractionation kit (Thermo Fisher Scientific, USA).
Flow Cytometry:Article Title: CtIP-Mediated Fork Protection Synergizes with BRCA1 to Suppress Genomic Instability upon DNA Replication Stress.
Article Snippet: Capan-1 ATCC Cat#HTB-79; RRID: CVCL_0237 H1299 DOX-inducible BRCA2 shRNA Madalena Tarsounas lab, Zimmer et al., 2016 N/A U2OS Flp-In T-REx Daniel Durocher lab, Al-Hakim et al., 2012 N/A PD20F (FANCD2-deficient) Fanconi Anemia Research Foundation (FARF), Murina et al., 2014 N/A PD20F complemented with FANCD2 WT Fanconi Anemia Research Foundation (FARF), Murina et al., 2014 N/A (Continued on next page) Molecular Cell 72, 1–15.e1–e6, November 1, 2018 e2 .. REAGENT or RESOURCE SOURCE IDENTIFIER Oligonucleotides Table S1 N/A N/A Recombinant DNA pEGFP-C1-CtIP (WT, T847A, S327A) Murina et al., 2014 N/A pEGFP-C1-CtIP N181A This paper N/A pcDNA5/FRT/TO-GFP-CtIP WT Ferretti et al., 2016 N/A pcDNA5/FRT/TO-GFP-CtIP (N181A, N289A/H290A) This paper N/A DR-GFP vector (pHPRT-DRGFP) Addgene #26476 N/A I-SceI expression plasmid (pCBASce) Addgene #26477 N/A DU46129, U6_gRNA MRC PPU Reagents and Services https://mrcppureagents.dundee.ac.uk Software and Algorithms GraphPad Prism7 for Mac OS X GraphPad Software http://www.graphpad.com/scientific- software/prism; RRID: SCR_002798 Fiji Department of Health and Human Services, NIDA https://fiji.sc/#; RRID: SCR_002285 Cell Profiler 2.2.0 Broad Institute http://cellprofiler.org; RRID: SCR_007358 FlowJo X FlowJo Software https://www.flowjo.com/; RRID: SCR_008520 FusionCapt Advance Solo 7 17.02 Vilber Lourmat http://www.vilber.de/ Olympus ScanR Image Analysis Software Version 2.5.1 OLYMPUS Image Analysis Software https://www.olympus-ims.com/pl/ microscope/software/ Spotfire TIBCO Software https://spotfire.tibco.com/; RRID: SCR_008858 Other BD FACSAria III BD Biosciences N/A Fusion Solo S Vilber N/A Leica DMI6000 Leica N/A BD LSR II Fortessa BD Biosciences N/A Attune NxT Flow Cytometer Thermo Fisher Scientific N/A CEF-DR II Variable Angle System Biorad N/A SpectraMax i3 Microplate Reader Molecular Devices N/A Olympus IX83 Olympus N/A Olympus ScanR Screening System Olympus N/A Faxitron Cabinet X-ray System Model RX-650 Faxitron N/A ..
Article Title: PALB2 deficiency may sensitize H3K27M-mutant pediatric HGG cells to BMN673/talazoparib
Article Snippet: .. The indicated EJ5 reporter cell lines were transfected with two plasmids, namely, the I-SceI expression vector pCBASce (Addgene #26477) and the DsRed expression vector (Addgene #54493), 48 h later, the percentage of GFP-positive cells among DsRed-positive cells was evaluated by imaging with a Carl Zeiss LSM 710 confocal fluorescence microscope and quantified via FACS on a BD AccuriTM C6 flow cytometer (BD Biosciences, USA) using BD AccuriTM software. .. Subcellular protein fractions were collected with a subcellular fractionation kit (Thermo Fisher Scientific, USA).
Transfection:
Article Title: PALB2 deficiency may sensitize H3K27M-mutant pediatric HGG cells to BMN673/talazoparib
Article Snippet: .. The indicated EJ5 reporter cell lines were transfected with two plasmids, namely, the I-SceI expression vector pCBASce (Addgene #26477) and the DsRed expression vector (Addgene #54493), 48 h later, the percentage of GFP-positive cells among DsRed-positive cells was evaluated by imaging with a Carl Zeiss LSM 710 confocal fluorescence microscope and quantified via FACS on a BD AccuriTM C6 flow cytometer (BD Biosciences, USA) using BD AccuriTM software. .. Subcellular protein fractions were collected with a subcellular fractionation kit (Thermo Fisher Scientific, USA).
Imaging:Article Title: PALB2 deficiency may sensitize H3K27M-mutant pediatric HGG cells to BMN673/talazoparib
Article Snippet: .. The indicated EJ5 reporter cell lines were transfected with two plasmids, namely, the I-SceI expression vector pCBASce (Addgene #26477) and the DsRed expression vector (Addgene #54493), 48 h later, the percentage of GFP-positive cells among DsRed-positive cells was evaluated by imaging with a Carl Zeiss LSM 710 confocal fluorescence microscope and quantified via FACS on a BD AccuriTM C6 flow cytometer (BD Biosciences, USA) using BD AccuriTM software. .. Subcellular protein fractions were collected with a subcellular fractionation kit (Thermo Fisher Scientific, USA).
Fluorescence:Article Title: PALB2 deficiency may sensitize H3K27M-mutant pediatric HGG cells to BMN673/talazoparib
Article Snippet: .. The indicated EJ5 reporter cell lines were transfected with two plasmids, namely, the I-SceI expression vector pCBASce (Addgene #26477) and the DsRed expression vector (Addgene #54493), 48 h later, the percentage of GFP-positive cells among DsRed-positive cells was evaluated by imaging with a Carl Zeiss LSM 710 confocal fluorescence microscope and quantified via FACS on a BD AccuriTM C6 flow cytometer (BD Biosciences, USA) using BD AccuriTM software. .. Subcellular protein fractions were collected with a subcellular fractionation kit (Thermo Fisher Scientific, USA).
FACS:Article Title: PALB2 deficiency may sensitize H3K27M-mutant pediatric HGG cells to BMN673/talazoparib
Article Snippet: .. The indicated EJ5 reporter cell lines were transfected with two plasmids, namely, the I-SceI expression vector pCBASce (Addgene #26477) and the DsRed expression vector (Addgene #54493), 48 h later, the percentage of GFP-positive cells among DsRed-positive cells was evaluated by imaging with a Carl Zeiss LSM 710 confocal fluorescence microscope and quantified via FACS on a BD AccuriTM C6 flow cytometer (BD Biosciences, USA) using BD AccuriTM software. .. Subcellular protein fractions were collected with a subcellular fractionation kit (Thermo Fisher Scientific, USA).
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