Infrared:Article Title: A Disease-Causing Single Amino Acid Deletion in the Coiled-Coil Domain of RAD50 Impairs MRE11 Complex Functions in Yeast and Humans
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Alexa Fluor 488 goat F[Ab0]2 Invitrogen A31556 Anti-53BP1 Novus Biological NB100-304; RRID: AB_10003037 Anti-vinculin Santa Cruz SC73614; RRID: AB_1131294 Anti-MRE11 Novus NB100-473; RRID: AB_10001780 Anti-RAD50 Genetex GTX 70228; RRID: AB_372854 Anti-NBS1 Novus NB100-143E1; RRID: AB_350080 Anti-CHK2 Santa Cruz SC17748; RRID: AB_627260 Goat anti-rabbit Ig IRDye 800CW Infrared Dye Li-Cor 926-32211; RRID: AB_621843 Anti-Phospho-KAP1 Bethyl S824 IHC 00073; RRID: AB_577234 Anti-RAD51 Millipore Calbiochem PC130; RRID: AB_2238184 Anti-RPA32 Bethyl A300-244A; RRID: AB_185548 Anti-BrdU Abcam clone BU1/75-ICR, Ab6326; RRID: AB_305426 Cy5 goat anti rat Ig Abcam AB6565; RRID: AB_955063 Cy3.5 goat anti mouse Ig Abcam AB6946; RRID: AB_955045 Anti-ssDNA antibodies Merck Millipore MAB3034; RRID: AB_94645 Anti-P-H3 Millipore 06-570; RRID: AB_310177 Anti-KU70 Thermo Scientific Pierce MA5-13110; RRID: AB_10976973 Anti-Phospho-NBS1 Cell Signaling 3001S; RRID: AB_10829154 Chemicals, Peptides, and Recombinant Proteins Triton X-100 Sigma 101958312 4’-6-diamidino-2-phenylindole dihydrochloride Thermo Scientific 62248 Phleomycin InvivoGen 11006-33-0 MMC Sigma M0503 Etoposide Sigma E1383 IdU Sigma I-7125 CldU Sigma C-6891 Fluorsave Calbiochem 345789 Bradford Bio-Rad 500-0006 Protease Inhibitor Roche 4693159001 Phosphatase inhibitor #1 Sigma P0044-5ML Phosphatase inhibitor #2 Sigma P5726-5ML PBS GIBCO 70011-36 PFA Sigma 1002491639 Glycine Euromedex 26-1286405C BSA Sigma A2153-100G Trizol Ambion 15596026 Tris Invitrogen 15504-020 EDTA Invitrogen 15575-038 Blocking buffer (TBS) Li-Cor 927-60001 Tween-20 MP Biochemicals 103168 Ethanol VWR 20821.330 IP Sigma P4864 Protéinase K Invitrogen P100005393 (Continued on next page) Cell Reports 33, 108559, December 29, 2020 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER b-Agarase New England Biolabs M0392S BrdU Becton Dickinson 550891 Critical Commercial Assays jetPRIME PolyPlus Transfection 114-07 FluorSave Calbiochem 345789 SureSelect Human All Exon Kit Agilent Technologies V5 SuperScript First-Strand Synthesis Kit Invitrogen 18064-014 QuikChange site-directed mutagenesis kit Agilent Technologies 200521 Experimental Models: Cell Lines Spodoptera frugiperda 9 (Sf9) cells Cejka lab N/A HT1080 Revy Lab N/A HT1080 RAD50+/ This paper N/A HT1080 RAD50-/del669-672 This paper N/A Primary fibroblast from healthy donor This paper N/A Primary fibroblast from patient P1 This paper N/A SV40-transformed hTERT-immortalized fibroblast from healthy donor This paper N/A SV40-transformed hTERT-immortalized fibroblast from patient P1 This paper N/A SV40-transformed hTERT-immortalized fibroblast from healthy donor transduced with pLVX-IRES-mCherry This paper N/A SV40-transformed hTERT-immortalized fibroblast from patient P1 transduced with pLVX-IRES-mCherry This paper N/A SV40-transformed hTERT-immortalized fibroblast from healthy donor transduced with pLVX-wtRAD50-IRES-mCherry This paper N/A SV40-transformed hTERT-immortalized fibroblast from patient P1 transduced with pLVX- wtRAD50-IRES-mCherry This paper N/A All S.cerevisiae strains used in this study are listed in Table S2 This paper N/A Experimental Models: Organisms/Strains All S.cerevisiae strains used in this study are listed in Table S2 This paper N/A Oligonucleotides codon-optimized oligonucleotides for recombinant protein: RAD50_DE1035_F (50- AACGAGGAGCT GAAGGAGGTGGAGGAGCGAAAGCAGCACCTG-30 This paper N/A Recombinant DNA PGK-H2BmCherry Addgene #21217 pX330-LMNA-gRNA1 Addgene #122507 pCR2.1 Clover-LMNA donor Addgene #122508 pLVX-EF1alpha-IRES-mCherry Takara Bio 631987 pLVX-EF1a-wtRAD50-IRES-mCherry This paper N/A pX330-hRAD50-gRNA This paper N/A pEGFPN1-NBS1 Lukas lab N/A pEGFP-NLS Zhang et al., 2016 N/A pLV-hTERT-IRES-hygro Addgene #85140 Software and Algorithms FlowJo FlowJo V10.1 GraphPad Prism version 8 GraphPad V6.0 (Continued on next page) e2 Cell Reports 33, 108559, December 29, 2020 Article ll OPEN ACCESS
Immunohistochemistry:Article Title: A Disease-Causing Single Amino Acid Deletion in the Coiled-Coil Domain of RAD50 Impairs MRE11 Complex Functions in Yeast and Humans
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Alexa Fluor 488 goat F[Ab0]2 Invitrogen A31556 Anti-53BP1 Novus Biological NB100-304; RRID: AB_10003037 Anti-vinculin Santa Cruz SC73614; RRID: AB_1131294 Anti-MRE11 Novus NB100-473; RRID: AB_10001780 Anti-RAD50 Genetex GTX 70228; RRID: AB_372854 Anti-NBS1 Novus NB100-143E1; RRID: AB_350080 Anti-CHK2 Santa Cruz SC17748; RRID: AB_627260 Goat anti-rabbit Ig IRDye 800CW Infrared Dye Li-Cor 926-32211; RRID: AB_621843 Anti-Phospho-KAP1 Bethyl S824 IHC 00073; RRID: AB_577234 Anti-RAD51 Millipore Calbiochem PC130; RRID: AB_2238184 Anti-RPA32 Bethyl A300-244A; RRID: AB_185548 Anti-BrdU Abcam clone BU1/75-ICR, Ab6326; RRID: AB_305426 Cy5 goat anti rat Ig Abcam AB6565; RRID: AB_955063 Cy3.5 goat anti mouse Ig Abcam AB6946; RRID: AB_955045 Anti-ssDNA antibodies Merck Millipore MAB3034; RRID: AB_94645 Anti-P-H3 Millipore 06-570; RRID: AB_310177 Anti-KU70 Thermo Scientific Pierce MA5-13110; RRID: AB_10976973 Anti-Phospho-NBS1 Cell Signaling 3001S; RRID: AB_10829154 Chemicals, Peptides, and Recombinant Proteins Triton X-100 Sigma 101958312 4’-6-diamidino-2-phenylindole dihydrochloride Thermo Scientific 62248 Phleomycin InvivoGen 11006-33-0 MMC Sigma M0503 Etoposide Sigma E1383 IdU Sigma I-7125 CldU Sigma C-6891 Fluorsave Calbiochem 345789 Bradford Bio-Rad 500-0006 Protease Inhibitor Roche 4693159001 Phosphatase inhibitor #1 Sigma P0044-5ML Phosphatase inhibitor #2 Sigma P5726-5ML PBS GIBCO 70011-36 PFA Sigma 1002491639 Glycine Euromedex 26-1286405C BSA Sigma A2153-100G Trizol Ambion 15596026 Tris Invitrogen 15504-020 EDTA Invitrogen 15575-038 Blocking buffer (TBS) Li-Cor 927-60001 Tween-20 MP Biochemicals 103168 Ethanol VWR 20821.330 IP Sigma P4864 Protéinase K Invitrogen P100005393 (Continued on next page) Cell Reports 33, 108559, December 29, 2020 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER b-Agarase New England Biolabs M0392S BrdU Becton Dickinson 550891 Critical Commercial Assays jetPRIME PolyPlus Transfection 114-07 FluorSave Calbiochem 345789 SureSelect Human All Exon Kit Agilent Technologies V5 SuperScript First-Strand Synthesis Kit Invitrogen 18064-014 QuikChange site-directed mutagenesis kit Agilent Technologies 200521 Experimental Models: Cell Lines Spodoptera frugiperda 9 (Sf9) cells Cejka lab N/A HT1080 Revy Lab N/A HT1080 RAD50+/ This paper N/A HT1080 RAD50-/del669-672 This paper N/A Primary fibroblast from healthy donor This paper N/A Primary fibroblast from patient P1 This paper N/A SV40-transformed hTERT-immortalized fibroblast from healthy donor This paper N/A SV40-transformed hTERT-immortalized fibroblast from patient P1 This paper N/A SV40-transformed hTERT-immortalized fibroblast from healthy donor transduced with pLVX-IRES-mCherry This paper N/A SV40-transformed hTERT-immortalized fibroblast from patient P1 transduced with pLVX-IRES-mCherry This paper N/A SV40-transformed hTERT-immortalized fibroblast from healthy donor transduced with pLVX-wtRAD50-IRES-mCherry This paper N/A SV40-transformed hTERT-immortalized fibroblast from patient P1 transduced with pLVX- wtRAD50-IRES-mCherry This paper N/A All S.cerevisiae strains used in this study are listed in Table S2 This paper N/A Experimental Models: Organisms/Strains All S.cerevisiae strains used in this study are listed in Table S2 This paper N/A Oligonucleotides codon-optimized oligonucleotides for recombinant protein: RAD50_DE1035_F (50- AACGAGGAGCT GAAGGAGGTGGAGGAGCGAAAGCAGCACCTG-30 This paper N/A Recombinant DNA PGK-H2BmCherry Addgene #21217 pX330-LMNA-gRNA1 Addgene #122507 pCR2.1 Clover-LMNA donor Addgene #122508 pLVX-EF1alpha-IRES-mCherry Takara Bio 631987 pLVX-EF1a-wtRAD50-IRES-mCherry This paper N/A pX330-hRAD50-gRNA This paper N/A pEGFPN1-NBS1 Lukas lab N/A pEGFP-NLS Zhang et al., 2016 N/A pLV-hTERT-IRES-hygro Addgene #85140 Software and Algorithms FlowJo FlowJo V10.1 GraphPad Prism version 8 GraphPad V6.0 (Continued on next page) e2 Cell Reports 33, 108559, December 29, 2020 Article ll OPEN ACCESS
Cell-Signaling:Article Title: A Disease-Causing Single Amino Acid Deletion in the Coiled-Coil Domain of RAD50 Impairs MRE11 Complex Functions in Yeast and Humans
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Alexa Fluor 488 goat F[Ab0]2 Invitrogen A31556 Anti-53BP1 Novus Biological NB100-304; RRID: AB_10003037 Anti-vinculin Santa Cruz SC73614; RRID: AB_1131294 Anti-MRE11 Novus NB100-473; RRID: AB_10001780 Anti-RAD50 Genetex GTX 70228; RRID: AB_372854 Anti-NBS1 Novus NB100-143E1; RRID: AB_350080 Anti-CHK2 Santa Cruz SC17748; RRID: AB_627260 Goat anti-rabbit Ig IRDye 800CW Infrared Dye Li-Cor 926-32211; RRID: AB_621843 Anti-Phospho-KAP1 Bethyl S824 IHC 00073; RRID: AB_577234 Anti-RAD51 Millipore Calbiochem PC130; RRID: AB_2238184 Anti-RPA32 Bethyl A300-244A; RRID: AB_185548 Anti-BrdU Abcam clone BU1/75-ICR, Ab6326; RRID: AB_305426 Cy5 goat anti rat Ig Abcam AB6565; RRID: AB_955063 Cy3.5 goat anti mouse Ig Abcam AB6946; RRID: AB_955045 Anti-ssDNA antibodies Merck Millipore MAB3034; RRID: AB_94645 Anti-P-H3 Millipore 06-570; RRID: AB_310177 Anti-KU70 Thermo Scientific Pierce MA5-13110; RRID: AB_10976973 Anti-Phospho-NBS1 Cell Signaling 3001S; RRID: AB_10829154 Chemicals, Peptides, and Recombinant Proteins Triton X-100 Sigma 101958312 4’-6-diamidino-2-phenylindole dihydrochloride Thermo Scientific 62248 Phleomycin InvivoGen 11006-33-0 MMC Sigma M0503 Etoposide Sigma E1383 IdU Sigma I-7125 CldU Sigma C-6891 Fluorsave Calbiochem 345789 Bradford Bio-Rad 500-0006 Protease Inhibitor Roche 4693159001 Phosphatase inhibitor #1 Sigma P0044-5ML Phosphatase inhibitor #2 Sigma P5726-5ML PBS GIBCO 70011-36 PFA Sigma 1002491639 Glycine Euromedex 26-1286405C BSA Sigma A2153-100G Trizol Ambion 15596026 Tris Invitrogen 15504-020 EDTA Invitrogen 15575-038 Blocking buffer (TBS) Li-Cor 927-60001 Tween-20 MP Biochemicals 103168 Ethanol VWR 20821.330 IP Sigma P4864 Protéinase K Invitrogen P100005393 (Continued on next page) Cell Reports 33, 108559, December 29, 2020 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER b-Agarase New England Biolabs M0392S BrdU Becton Dickinson 550891 Critical Commercial Assays jetPRIME PolyPlus Transfection 114-07 FluorSave Calbiochem 345789 SureSelect Human All Exon Kit Agilent Technologies V5 SuperScript First-Strand Synthesis Kit Invitrogen 18064-014 QuikChange site-directed mutagenesis kit Agilent Technologies 200521 Experimental Models: Cell Lines Spodoptera frugiperda 9 (Sf9) cells Cejka lab N/A HT1080 Revy Lab N/A HT1080 RAD50+/ This paper N/A HT1080 RAD50-/del669-672 This paper N/A Primary fibroblast from healthy donor This paper N/A Primary fibroblast from patient P1 This paper N/A SV40-transformed hTERT-immortalized fibroblast from healthy donor This paper N/A SV40-transformed hTERT-immortalized fibroblast from patient P1 This paper N/A SV40-transformed hTERT-immortalized fibroblast from healthy donor transduced with pLVX-IRES-mCherry This paper N/A SV40-transformed hTERT-immortalized fibroblast from patient P1 transduced with pLVX-IRES-mCherry This paper N/A SV40-transformed hTERT-immortalized fibroblast from healthy donor transduced with pLVX-wtRAD50-IRES-mCherry This paper N/A SV40-transformed hTERT-immortalized fibroblast from patient P1 transduced with pLVX- wtRAD50-IRES-mCherry This paper N/A All S.cerevisiae strains used in this study are listed in Table S2 This paper N/A Experimental Models: Organisms/Strains All S.cerevisiae strains used in this study are listed in Table S2 This paper N/A Oligonucleotides codon-optimized oligonucleotides for recombinant protein: RAD50_DE1035_F (50- AACGAGGAGCT GAAGGAGGTGGAGGAGCGAAAGCAGCACCTG-30 This paper N/A Recombinant DNA PGK-H2BmCherry Addgene #21217 pX330-LMNA-gRNA1 Addgene #122507 pCR2.1 Clover-LMNA donor Addgene #122508 pLVX-EF1alpha-IRES-mCherry Takara Bio 631987 pLVX-EF1a-wtRAD50-IRES-mCherry This paper N/A pX330-hRAD50-gRNA This paper N/A pEGFPN1-NBS1 Lukas lab N/A pEGFP-NLS Zhang et al., 2016 N/A pLV-hTERT-IRES-hygro Addgene #85140 Software and Algorithms FlowJo FlowJo V10.1 GraphPad Prism version 8 GraphPad V6.0 (Continued on next page) e2 Cell Reports 33, 108559, December 29, 2020 Article ll OPEN ACCESS
Recombinant:Article Title: A Disease-Causing Single Amino Acid Deletion in the Coiled-Coil Domain of RAD50 Impairs MRE11 Complex Functions in Yeast and Humans
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Alexa Fluor 488 goat F[Ab0]2 Invitrogen A31556 Anti-53BP1 Novus Biological NB100-304; RRID: AB_10003037 Anti-vinculin Santa Cruz SC73614; RRID: AB_1131294 Anti-MRE11 Novus NB100-473; RRID: AB_10001780 Anti-RAD50 Genetex GTX 70228; RRID: AB_372854 Anti-NBS1 Novus NB100-143E1; RRID: AB_350080 Anti-CHK2 Santa Cruz SC17748; RRID: AB_627260 Goat anti-rabbit Ig IRDye 800CW Infrared Dye Li-Cor 926-32211; RRID: AB_621843 Anti-Phospho-KAP1 Bethyl S824 IHC 00073; RRID: AB_577234 Anti-RAD51 Millipore Calbiochem PC130; RRID: AB_2238184 Anti-RPA32 Bethyl A300-244A; RRID: AB_185548 Anti-BrdU Abcam clone BU1/75-ICR, Ab6326; RRID: AB_305426 Cy5 goat anti rat Ig Abcam AB6565; RRID: AB_955063 Cy3.5 goat anti mouse Ig Abcam AB6946; RRID: AB_955045 Anti-ssDNA antibodies Merck Millipore MAB3034; RRID: AB_94645 Anti-P-H3 Millipore 06-570; RRID: AB_310177 Anti-KU70 Thermo Scientific Pierce MA5-13110; RRID: AB_10976973 Anti-Phospho-NBS1 Cell Signaling 3001S; RRID: AB_10829154 Chemicals, Peptides, and Recombinant Proteins Triton X-100 Sigma 101958312 4’-6-diamidino-2-phenylindole dihydrochloride Thermo Scientific 62248 Phleomycin InvivoGen 11006-33-0 MMC Sigma M0503 Etoposide Sigma E1383 IdU Sigma I-7125 CldU Sigma C-6891 Fluorsave Calbiochem 345789 Bradford Bio-Rad 500-0006 Protease Inhibitor Roche 4693159001 Phosphatase inhibitor #1 Sigma P0044-5ML Phosphatase inhibitor #2 Sigma P5726-5ML PBS GIBCO 70011-36 PFA Sigma 1002491639 Glycine Euromedex 26-1286405C BSA Sigma A2153-100G Trizol Ambion 15596026 Tris Invitrogen 15504-020 EDTA Invitrogen 15575-038 Blocking buffer (TBS) Li-Cor 927-60001 Tween-20 MP Biochemicals 103168 Ethanol VWR 20821.330 IP Sigma P4864 Protéinase K Invitrogen P100005393 (Continued on next page) Cell Reports 33, 108559, December 29, 2020 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER b-Agarase New England Biolabs M0392S BrdU Becton Dickinson 550891 Critical Commercial Assays jetPRIME PolyPlus Transfection 114-07 FluorSave Calbiochem 345789 SureSelect Human All Exon Kit Agilent Technologies V5 SuperScript First-Strand Synthesis Kit Invitrogen 18064-014 QuikChange site-directed mutagenesis kit Agilent Technologies 200521 Experimental Models: Cell Lines Spodoptera frugiperda 9 (Sf9) cells Cejka lab N/A HT1080 Revy Lab N/A HT1080 RAD50+/ This paper N/A HT1080 RAD50-/del669-672 This paper N/A Primary fibroblast from healthy donor This paper N/A Primary fibroblast from patient P1 This paper N/A SV40-transformed hTERT-immortalized fibroblast from healthy donor This paper N/A SV40-transformed hTERT-immortalized fibroblast from patient P1 This paper N/A SV40-transformed hTERT-immortalized fibroblast from healthy donor transduced with pLVX-IRES-mCherry This paper N/A SV40-transformed hTERT-immortalized fibroblast from patient P1 transduced with pLVX-IRES-mCherry This paper N/A SV40-transformed hTERT-immortalized fibroblast from healthy donor transduced with pLVX-wtRAD50-IRES-mCherry This paper N/A SV40-transformed hTERT-immortalized fibroblast from patient P1 transduced with pLVX- wtRAD50-IRES-mCherry This paper N/A All S.cerevisiae strains used in this study are listed in Table S2 This paper N/A Experimental Models: Organisms/Strains All S.cerevisiae strains used in this study are listed in Table S2 This paper N/A Oligonucleotides codon-optimized oligonucleotides for recombinant protein: RAD50_DE1035_F (50- AACGAGGAGCT GAAGGAGGTGGAGGAGCGAAAGCAGCACCTG-30 This paper N/A Recombinant DNA PGK-H2BmCherry Addgene #21217 pX330-LMNA-gRNA1 Addgene #122507 pCR2.1 Clover-LMNA donor Addgene #122508 pLVX-EF1alpha-IRES-mCherry Takara Bio 631987 pLVX-EF1a-wtRAD50-IRES-mCherry This paper N/A pX330-hRAD50-gRNA This paper N/A pEGFPN1-NBS1 Lukas lab N/A pEGFP-NLS Zhang et al., 2016 N/A pLV-hTERT-IRES-hygro Addgene #85140 Software and Algorithms FlowJo FlowJo V10.1 GraphPad Prism version 8 GraphPad V6.0 (Continued on next page) e2 Cell Reports 33, 108559, December 29, 2020 Article ll OPEN ACCESS
Fluorsave:Article Title: A Disease-Causing Single Amino Acid Deletion in the Coiled-Coil Domain of RAD50 Impairs MRE11 Complex Functions in Yeast and Humans
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Alexa Fluor 488 goat F[Ab0]2 Invitrogen A31556 Anti-53BP1 Novus Biological NB100-304; RRID: AB_10003037 Anti-vinculin Santa Cruz SC73614; RRID: AB_1131294 Anti-MRE11 Novus NB100-473; RRID: AB_10001780 Anti-RAD50 Genetex GTX 70228; RRID: AB_372854 Anti-NBS1 Novus NB100-143E1; RRID: AB_350080 Anti-CHK2 Santa Cruz SC17748; RRID: AB_627260 Goat anti-rabbit Ig IRDye 800CW Infrared Dye Li-Cor 926-32211; RRID: AB_621843 Anti-Phospho-KAP1 Bethyl S824 IHC 00073; RRID: AB_577234 Anti-RAD51 Millipore Calbiochem PC130; RRID: AB_2238184 Anti-RPA32 Bethyl A300-244A; RRID: AB_185548 Anti-BrdU Abcam clone BU1/75-ICR, Ab6326; RRID: AB_305426 Cy5 goat anti rat Ig Abcam AB6565; RRID: AB_955063 Cy3.5 goat anti mouse Ig Abcam AB6946; RRID: AB_955045 Anti-ssDNA antibodies Merck Millipore MAB3034; RRID: AB_94645 Anti-P-H3 Millipore 06-570; RRID: AB_310177 Anti-KU70 Thermo Scientific Pierce MA5-13110; RRID: AB_10976973 Anti-Phospho-NBS1 Cell Signaling 3001S; RRID: AB_10829154 Chemicals, Peptides, and Recombinant Proteins Triton X-100 Sigma 101958312 4’-6-diamidino-2-phenylindole dihydrochloride Thermo Scientific 62248 Phleomycin InvivoGen 11006-33-0 MMC Sigma M0503 Etoposide Sigma E1383 IdU Sigma I-7125 CldU Sigma C-6891 Fluorsave Calbiochem 345789 Bradford Bio-Rad 500-0006 Protease Inhibitor Roche 4693159001 Phosphatase inhibitor #1 Sigma P0044-5ML Phosphatase inhibitor #2 Sigma P5726-5ML PBS GIBCO 70011-36 PFA Sigma 1002491639 Glycine Euromedex 26-1286405C BSA Sigma A2153-100G Trizol Ambion 15596026 Tris Invitrogen 15504-020 EDTA Invitrogen 15575-038 Blocking buffer (TBS) Li-Cor 927-60001 Tween-20 MP Biochemicals 103168 Ethanol VWR 20821.330 IP Sigma P4864 Protéinase K Invitrogen P100005393 (Continued on next page) Cell Reports 33, 108559, December 29, 2020 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER b-Agarase New England Biolabs M0392S BrdU Becton Dickinson 550891 Critical Commercial Assays jetPRIME PolyPlus Transfection 114-07 FluorSave Calbiochem 345789 SureSelect Human All Exon Kit Agilent Technologies V5 SuperScript First-Strand Synthesis Kit Invitrogen 18064-014 QuikChange site-directed mutagenesis kit Agilent Technologies 200521 Experimental Models: Cell Lines Spodoptera frugiperda 9 (Sf9) cells Cejka lab N/A HT1080 Revy Lab N/A HT1080 RAD50+/ This paper N/A HT1080 RAD50-/del669-672 This paper N/A Primary fibroblast from healthy donor This paper N/A Primary fibroblast from patient P1 This paper N/A SV40-transformed hTERT-immortalized fibroblast from healthy donor This paper N/A SV40-transformed hTERT-immortalized fibroblast from patient P1 This paper N/A SV40-transformed hTERT-immortalized fibroblast from healthy donor transduced with pLVX-IRES-mCherry This paper N/A SV40-transformed hTERT-immortalized fibroblast from patient P1 transduced with pLVX-IRES-mCherry This paper N/A SV40-transformed hTERT-immortalized fibroblast from healthy donor transduced with pLVX-wtRAD50-IRES-mCherry This paper N/A SV40-transformed hTERT-immortalized fibroblast from patient P1 transduced with pLVX- wtRAD50-IRES-mCherry This paper N/A All S.cerevisiae strains used in this study are listed in Table S2 This paper N/A Experimental Models: Organisms/Strains All S.cerevisiae strains used in this study are listed in Table S2 This paper N/A Oligonucleotides codon-optimized oligonucleotides for recombinant protein: RAD50_DE1035_F (50- AACGAGGAGCT GAAGGAGGTGGAGGAGCGAAAGCAGCACCTG-30 This paper N/A Recombinant DNA PGK-H2BmCherry Addgene #21217 pX330-LMNA-gRNA1 Addgene #122507 pCR2.1 Clover-LMNA donor Addgene #122508 pLVX-EF1alpha-IRES-mCherry Takara Bio 631987 pLVX-EF1a-wtRAD50-IRES-mCherry This paper N/A pX330-hRAD50-gRNA This paper N/A pEGFPN1-NBS1 Lukas lab N/A pEGFP-NLS Zhang et al., 2016 N/A pLV-hTERT-IRES-hygro Addgene #85140 Software and Algorithms FlowJo FlowJo V10.1 GraphPad Prism version 8 GraphPad V6.0 (Continued on next page) e2 Cell Reports 33, 108559, December 29, 2020 Article ll OPEN ACCESS
Protease Inhibitor:Article Title: A Disease-Causing Single Amino Acid Deletion in the Coiled-Coil Domain of RAD50 Impairs MRE11 Complex Functions in Yeast and Humans
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Alexa Fluor 488 goat F[Ab0]2 Invitrogen A31556 Anti-53BP1 Novus Biological NB100-304; RRID: AB_10003037 Anti-vinculin Santa Cruz SC73614; RRID: AB_1131294 Anti-MRE11 Novus NB100-473; RRID: AB_10001780 Anti-RAD50 Genetex GTX 70228; RRID: AB_372854 Anti-NBS1 Novus NB100-143E1; RRID: AB_350080 Anti-CHK2 Santa Cruz SC17748; RRID: AB_627260 Goat anti-rabbit Ig IRDye 800CW Infrared Dye Li-Cor 926-32211; RRID: AB_621843 Anti-Phospho-KAP1 Bethyl S824 IHC 00073; RRID: AB_577234 Anti-RAD51 Millipore Calbiochem PC130; RRID: AB_2238184 Anti-RPA32 Bethyl A300-244A; RRID: AB_185548 Anti-BrdU Abcam clone BU1/75-ICR, Ab6326; RRID: AB_305426 Cy5 goat anti rat Ig Abcam AB6565; RRID: AB_955063 Cy3.5 goat anti mouse Ig Abcam AB6946; RRID: AB_955045 Anti-ssDNA antibodies Merck Millipore MAB3034; RRID: AB_94645 Anti-P-H3 Millipore 06-570; RRID: AB_310177 Anti-KU70 Thermo Scientific Pierce MA5-13110; RRID: AB_10976973 Anti-Phospho-NBS1 Cell Signaling 3001S; RRID: AB_10829154 Chemicals, Peptides, and Recombinant Proteins Triton X-100 Sigma 101958312 4’-6-diamidino-2-phenylindole dihydrochloride Thermo Scientific 62248 Phleomycin InvivoGen 11006-33-0 MMC Sigma M0503 Etoposide Sigma E1383 IdU Sigma I-7125 CldU Sigma C-6891 Fluorsave Calbiochem 345789 Bradford Bio-Rad 500-0006 Protease Inhibitor Roche 4693159001 Phosphatase inhibitor #1 Sigma P0044-5ML Phosphatase inhibitor #2 Sigma P5726-5ML PBS GIBCO 70011-36 PFA Sigma 1002491639 Glycine Euromedex 26-1286405C BSA Sigma A2153-100G Trizol Ambion 15596026 Tris Invitrogen 15504-020 EDTA Invitrogen 15575-038 Blocking buffer (TBS) Li-Cor 927-60001 Tween-20 MP Biochemicals 103168 Ethanol VWR 20821.330 IP Sigma P4864 Protéinase K Invitrogen P100005393 (Continued on next page) Cell Reports 33, 108559, December 29, 2020 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER b-Agarase New England Biolabs M0392S BrdU Becton Dickinson 550891 Critical Commercial Assays jetPRIME PolyPlus Transfection 114-07 FluorSave Calbiochem 345789 SureSelect Human All Exon Kit Agilent Technologies V5 SuperScript First-Strand Synthesis Kit Invitrogen 18064-014 QuikChange site-directed mutagenesis kit Agilent Technologies 200521 Experimental Models: Cell Lines Spodoptera frugiperda 9 (Sf9) cells Cejka lab N/A HT1080 Revy Lab N/A HT1080 RAD50+/ This paper N/A HT1080 RAD50-/del669-672 This paper N/A Primary fibroblast from healthy donor This paper N/A Primary fibroblast from patient P1 This paper N/A SV40-transformed hTERT-immortalized fibroblast from healthy donor This paper N/A SV40-transformed hTERT-immortalized fibroblast from patient P1 This paper N/A SV40-transformed hTERT-immortalized fibroblast from healthy donor transduced with pLVX-IRES-mCherry This paper N/A SV40-transformed hTERT-immortalized fibroblast from patient P1 transduced with pLVX-IRES-mCherry This paper N/A SV40-transformed hTERT-immortalized fibroblast from healthy donor transduced with pLVX-wtRAD50-IRES-mCherry This paper N/A SV40-transformed hTERT-immortalized fibroblast from patient P1 transduced with pLVX- wtRAD50-IRES-mCherry This paper N/A All S.cerevisiae strains used in this study are listed in Table S2 This paper N/A Experimental Models: Organisms/Strains All S.cerevisiae strains used in this study are listed in Table S2 This paper N/A Oligonucleotides codon-optimized oligonucleotides for recombinant protein: RAD50_DE1035_F (50- AACGAGGAGCT GAAGGAGGTGGAGGAGCGAAAGCAGCACCTG-30 This paper N/A Recombinant DNA PGK-H2BmCherry Addgene #21217 pX330-LMNA-gRNA1 Addgene #122507 pCR2.1 Clover-LMNA donor Addgene #122508 pLVX-EF1alpha-IRES-mCherry Takara Bio 631987 pLVX-EF1a-wtRAD50-IRES-mCherry This paper N/A pX330-hRAD50-gRNA This paper N/A pEGFPN1-NBS1 Lukas lab N/A pEGFP-NLS Zhang et al., 2016 N/A pLV-hTERT-IRES-hygro Addgene #85140 Software and Algorithms FlowJo FlowJo V10.1 GraphPad Prism version 8 GraphPad V6.0 (Continued on next page) e2 Cell Reports 33, 108559, December 29, 2020 Article ll OPEN ACCESS
Blocking Assay:Article Title: A Disease-Causing Single Amino Acid Deletion in the Coiled-Coil Domain of RAD50 Impairs MRE11 Complex Functions in Yeast and Humans
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Alexa Fluor 488 goat F[Ab0]2 Invitrogen A31556 Anti-53BP1 Novus Biological NB100-304; RRID: AB_10003037 Anti-vinculin Santa Cruz SC73614; RRID: AB_1131294 Anti-MRE11 Novus NB100-473; RRID: AB_10001780 Anti-RAD50 Genetex GTX 70228; RRID: AB_372854 Anti-NBS1 Novus NB100-143E1; RRID: AB_350080 Anti-CHK2 Santa Cruz SC17748; RRID: AB_627260 Goat anti-rabbit Ig IRDye 800CW Infrared Dye Li-Cor 926-32211; RRID: AB_621843 Anti-Phospho-KAP1 Bethyl S824 IHC 00073; RRID: AB_577234 Anti-RAD51 Millipore Calbiochem PC130; RRID: AB_2238184 Anti-RPA32 Bethyl A300-244A; RRID: AB_185548 Anti-BrdU Abcam clone BU1/75-ICR, Ab6326; RRID: AB_305426 Cy5 goat anti rat Ig Abcam AB6565; RRID: AB_955063 Cy3.5 goat anti mouse Ig Abcam AB6946; RRID: AB_955045 Anti-ssDNA antibodies Merck Millipore MAB3034; RRID: AB_94645 Anti-P-H3 Millipore 06-570; RRID: AB_310177 Anti-KU70 Thermo Scientific Pierce MA5-13110; RRID: AB_10976973 Anti-Phospho-NBS1 Cell Signaling 3001S; RRID: AB_10829154 Chemicals, Peptides, and Recombinant Proteins Triton X-100 Sigma 101958312 4’-6-diamidino-2-phenylindole dihydrochloride Thermo Scientific 62248 Phleomycin InvivoGen 11006-33-0 MMC Sigma M0503 Etoposide Sigma E1383 IdU Sigma I-7125 CldU Sigma C-6891 Fluorsave Calbiochem 345789 Bradford Bio-Rad 500-0006 Protease Inhibitor Roche 4693159001 Phosphatase inhibitor #1 Sigma P0044-5ML Phosphatase inhibitor #2 Sigma P5726-5ML PBS GIBCO 70011-36 PFA Sigma 1002491639 Glycine Euromedex 26-1286405C BSA Sigma A2153-100G Trizol Ambion 15596026 Tris Invitrogen 15504-020 EDTA Invitrogen 15575-038 Blocking buffer (TBS) Li-Cor 927-60001 Tween-20 MP Biochemicals 103168 Ethanol VWR 20821.330 IP Sigma P4864 Protéinase K Invitrogen P100005393 (Continued on next page) Cell Reports 33, 108559, December 29, 2020 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER b-Agarase New England Biolabs M0392S BrdU Becton Dickinson 550891 Critical Commercial Assays jetPRIME PolyPlus Transfection 114-07 FluorSave Calbiochem 345789 SureSelect Human All Exon Kit Agilent Technologies V5 SuperScript First-Strand Synthesis Kit Invitrogen 18064-014 QuikChange site-directed mutagenesis kit Agilent Technologies 200521 Experimental Models: Cell Lines Spodoptera frugiperda 9 (Sf9) cells Cejka lab N/A HT1080 Revy Lab N/A HT1080 RAD50+/ This paper N/A HT1080 RAD50-/del669-672 This paper N/A Primary fibroblast from healthy donor This paper N/A Primary fibroblast from patient P1 This paper N/A SV40-transformed hTERT-immortalized fibroblast from healthy donor This paper N/A SV40-transformed hTERT-immortalized fibroblast from patient P1 This paper N/A SV40-transformed hTERT-immortalized fibroblast from healthy donor transduced with pLVX-IRES-mCherry This paper N/A SV40-transformed hTERT-immortalized fibroblast from patient P1 transduced with pLVX-IRES-mCherry This paper N/A SV40-transformed hTERT-immortalized fibroblast from healthy donor transduced with pLVX-wtRAD50-IRES-mCherry This paper N/A SV40-transformed hTERT-immortalized fibroblast from patient P1 transduced with pLVX- wtRAD50-IRES-mCherry This paper N/A All S.cerevisiae strains used in this study are listed in Table S2 This paper N/A Experimental Models: Organisms/Strains All S.cerevisiae strains used in this study are listed in Table S2 This paper N/A Oligonucleotides codon-optimized oligonucleotides for recombinant protein: RAD50_DE1035_F (50- AACGAGGAGCT GAAGGAGGTGGAGGAGCGAAAGCAGCACCTG-30 This paper N/A Recombinant DNA PGK-H2BmCherry Addgene #21217 pX330-LMNA-gRNA1 Addgene #122507 pCR2.1 Clover-LMNA donor Addgene #122508 pLVX-EF1alpha-IRES-mCherry Takara Bio 631987 pLVX-EF1a-wtRAD50-IRES-mCherry This paper N/A pX330-hRAD50-gRNA This paper N/A pEGFPN1-NBS1 Lukas lab N/A pEGFP-NLS Zhang et al., 2016 N/A pLV-hTERT-IRES-hygro Addgene #85140 Software and Algorithms FlowJo FlowJo V10.1 GraphPad Prism version 8 GraphPad V6.0 (Continued on next page) e2 Cell Reports 33, 108559, December 29, 2020 Article ll OPEN ACCESS
Transcranial Magnetic Stimulation:Article Title: A Disease-Causing Single Amino Acid Deletion in the Coiled-Coil Domain of RAD50 Impairs MRE11 Complex Functions in Yeast and Humans
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Alexa Fluor 488 goat F[Ab0]2 Invitrogen A31556 Anti-53BP1 Novus Biological NB100-304; RRID: AB_10003037 Anti-vinculin Santa Cruz SC73614; RRID: AB_1131294 Anti-MRE11 Novus NB100-473; RRID: AB_10001780 Anti-RAD50 Genetex GTX 70228; RRID: AB_372854 Anti-NBS1 Novus NB100-143E1; RRID: AB_350080 Anti-CHK2 Santa Cruz SC17748; RRID: AB_627260 Goat anti-rabbit Ig IRDye 800CW Infrared Dye Li-Cor 926-32211; RRID: AB_621843 Anti-Phospho-KAP1 Bethyl S824 IHC 00073; RRID: AB_577234 Anti-RAD51 Millipore Calbiochem PC130; RRID: AB_2238184 Anti-RPA32 Bethyl A300-244A; RRID: AB_185548 Anti-BrdU Abcam clone BU1/75-ICR, Ab6326; RRID: AB_305426 Cy5 goat anti rat Ig Abcam AB6565; RRID: AB_955063 Cy3.5 goat anti mouse Ig Abcam AB6946; RRID: AB_955045 Anti-ssDNA antibodies Merck Millipore MAB3034; RRID: AB_94645 Anti-P-H3 Millipore 06-570; RRID: AB_310177 Anti-KU70 Thermo Scientific Pierce MA5-13110; RRID: AB_10976973 Anti-Phospho-NBS1 Cell Signaling 3001S; RRID: AB_10829154 Chemicals, Peptides, and Recombinant Proteins Triton X-100 Sigma 101958312 4’-6-diamidino-2-phenylindole dihydrochloride Thermo Scientific 62248 Phleomycin InvivoGen 11006-33-0 MMC Sigma M0503 Etoposide Sigma E1383 IdU Sigma I-7125 CldU Sigma C-6891 Fluorsave Calbiochem 345789 Bradford Bio-Rad 500-0006 Protease Inhibitor Roche 4693159001 Phosphatase inhibitor #1 Sigma P0044-5ML Phosphatase inhibitor #2 Sigma P5726-5ML PBS GIBCO 70011-36 PFA Sigma 1002491639 Glycine Euromedex 26-1286405C BSA Sigma A2153-100G Trizol Ambion 15596026 Tris Invitrogen 15504-020 EDTA Invitrogen 15575-038 Blocking buffer (TBS) Li-Cor 927-60001 Tween-20 MP Biochemicals 103168 Ethanol VWR 20821.330 IP Sigma P4864 Protéinase K Invitrogen P100005393 (Continued on next page) Cell Reports 33, 108559, December 29, 2020 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER b-Agarase New England Biolabs M0392S BrdU Becton Dickinson 550891 Critical Commercial Assays jetPRIME PolyPlus Transfection 114-07 FluorSave Calbiochem 345789 SureSelect Human All Exon Kit Agilent Technologies V5 SuperScript First-Strand Synthesis Kit Invitrogen 18064-014 QuikChange site-directed mutagenesis kit Agilent Technologies 200521 Experimental Models: Cell Lines Spodoptera frugiperda 9 (Sf9) cells Cejka lab N/A HT1080 Revy Lab N/A HT1080 RAD50+/ This paper N/A HT1080 RAD50-/del669-672 This paper N/A Primary fibroblast from healthy donor This paper N/A Primary fibroblast from patient P1 This paper N/A SV40-transformed hTERT-immortalized fibroblast from healthy donor This paper N/A SV40-transformed hTERT-immortalized fibroblast from patient P1 This paper N/A SV40-transformed hTERT-immortalized fibroblast from healthy donor transduced with pLVX-IRES-mCherry This paper N/A SV40-transformed hTERT-immortalized fibroblast from patient P1 transduced with pLVX-IRES-mCherry This paper N/A SV40-transformed hTERT-immortalized fibroblast from healthy donor transduced with pLVX-wtRAD50-IRES-mCherry This paper N/A SV40-transformed hTERT-immortalized fibroblast from patient P1 transduced with pLVX- wtRAD50-IRES-mCherry This paper N/A All S.cerevisiae strains used in this study are listed in Table S2 This paper N/A Experimental Models: Organisms/Strains All S.cerevisiae strains used in this study are listed in Table S2 This paper N/A Oligonucleotides codon-optimized oligonucleotides for recombinant protein: RAD50_DE1035_F (50- AACGAGGAGCT GAAGGAGGTGGAGGAGCGAAAGCAGCACCTG-30 This paper N/A Recombinant DNA PGK-H2BmCherry Addgene #21217 pX330-LMNA-gRNA1 Addgene #122507 pCR2.1 Clover-LMNA donor Addgene #122508 pLVX-EF1alpha-IRES-mCherry Takara Bio 631987 pLVX-EF1a-wtRAD50-IRES-mCherry This paper N/A pX330-hRAD50-gRNA This paper N/A pEGFPN1-NBS1 Lukas lab N/A pEGFP-NLS Zhang et al., 2016 N/A pLV-hTERT-IRES-hygro Addgene #85140 Software and Algorithms FlowJo FlowJo V10.1 GraphPad Prism version 8 GraphPad V6.0 (Continued on next page) e2 Cell Reports 33, 108559, December 29, 2020 Article ll OPEN ACCESS
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