other:
Incubation:Article Title: Telomere occupancy by TRF2 is altered by KIT mutations and correlates with mastocytosis regression.
Article Snippet: .. Primary anti-KIT (polyclonal, R&D, AF332, dilution 1/20), anti-TRF2 (4A794.15, Imgenex, IMG124A, dilution 1/20), anti-p-TRF2 (Thr188) (Polyclonal, ThermoFisher Scientific, PA5-105937, dilution 1/200) were incubated one hour at room temperature, secondary antibody donkey anti-IgG goat AF488 (Invitrogen, 1/200) was incubated 30min at room temperature. ..
Article Title: Telomere occupancy by TRF2 is altered by KIT mutations and correlates with mastocytosis regression
Article Snippet: .. Primary anti-KIT (polyclonal, R&D, AF332, dilution 1/20), anti-TRF2 (4A794.15, Imgenex, IMG-124A, dilution 1/20), anti-p-TRF2 (Thr188) (Polyclonal, ThermoFisher Scientific, PA5-105937, dilution 1/200) were incubated one hour at room temperature, secondary antibody donkey anti-IgG goat AF488 (Invitrogen, 1/200) was incubated 30 min at room temperature. ..
Virus:Article Title: TRF1 and TRF2 form distinct shelterin subcomplexes at telomeres.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse monoclonal anti-TERF2 Novus CatNB100-56506; RRID: AB_839054 Rabbit polyclonal anti-TERF2 Novus Cat#NB110-57130; RRID: AB_844199 Rabbit polyclonal anti-TERF2IP Novus Cat#NB100-292; RRID: AB_10000825 Rabbit polyclonal anti-TPP1 Bethyl Cat#A303-069A; RRID: AB_10890369 Rabbit polyclonal anti-POT1 Proteintech Cat#10581-1-AP; RRID: AB_2284057 Rabbit anti-TERF1 Jan Karlseder N/A Mouse monoclonal anti-FLAG (HRP) Sigma-Aldrich Cat#A8592; RRID: AB_439702 Rabbit polyclonal anti-53BP1 Novus Cat#NB100-304; RRID: AB_10003037 Goat polyclonal anti-rabbit (Cy3) Invitrogen Cat#A10520; RRID: AB_10563288 Goat polyclonal anti-rabbit (HRP) Invitrogen Cat#31460; RRID: AB_228341 Goat polyclonal anti-mouse (HRP) Invitrogen Cat#31439; RRID: AB_228292 Bacterial and virus strains One Shot BL21(DE3) Chemically Competent E. coli Invitrogen Cat#C600003 TOP10 Chemically Competent E. coli Invitrogen Cat#C404006 Chemicals, peptides, and recombinant proteins TEV protease New England Biolabs Cat#P8112 Lipofectamine 2000 Invitrogen Cat#11668500 HaloPROTAC-E AOBIOUS Inc. Cat#AOB13155 Ouabain Sigma-Aldrich Cat#O3125 Thymidine Sigma-Aldrich Cat#T1895 PNA TelC-Alexa488 PNA Bio Cat#F1004 Janelia Fluor JFX650 HaloTag Ligand Grimm et al. 78 ; Promega Cat#HT1070 Janelia Fluor 657 HaloTag Ligand Grimm et al. 79 N/A Janelia Fluor 549 HaloTag Ligand Grimm et al. 80 ; Promega Cat#HT1020 Hoechst 33342 Thermo Scientific Cat#62249 Prolong Diamond Antifade Mountant Invitrogen Cat#P36970 Super Nuclease Sino Biological Cat#SSNP01 ANTI-FLAG M2 Affinity Gel Millipore Cat#A2220 3xFLAG Peptide Sigma-Aldrich Cat#F4799 Ni Sepharose 6 Fast Flow histidine-tagged protein purification resin Cytiva Cat#17531801 Critical commercial assays FxCycle PI/RNase Staining Solution Invitrogen Cat#F10797 SuperSignal West Femto Maximum Sensitivity Substrate Thermo Scientific Cat#34094 4–15% Mini-PROTEAN TGX Stain-Free Protein Gels Bio-Rad Cat#4568083 4–15% Criterion TGX Stain-Free Protein Gel Bio-Rad Cat#5678084 Trans-Blot Turbo RTA Mini 0.2 μm Nitrocellulose Transfer Kit Bio-Rad Cat#1704270 Deposited data All generated plasmids will be deposited at Addgene N/A (Continued on next page) 16 Cell Reports 44, 116178, September 23, 2025 .. REAGENT or RESOURCE SOURCE IDENTIFIER Photobleaching and residence time analysis algorithms This study https://github.com/SchmidtLabMSU/ ShelterinAnalysis Experimental models: Cell lines Human: HeLa EM2-11ht Weidenfeld et al. 50 RRID: CVCL_WN71 Human: HeLa EM2-11ht, FLAG-HaloTag-TRF1 This study N/A Human: HeLa EM2-11ht, FLAG-HaloTag-TRF2 This study N/A Human: HeLa EM2-11ht, FLAG-HaloTag-TIN2 This study N/A Human: HeLa EM2-11ht, FLAG-HaloTag-POT1 This study N/A Human: HeLa EM2-11ht, FLAG-HaloTag-TRF2, RAP1-KO This study N/A Human: HeLa 1.3 Takai et al. 48 RRID: CVCL_B5DA Human: HeLa 1.3, FLAG-HaloTag-TRF2 This study N/A Human: HeLa 1.3, FLAG-HaloTag-TIN2 This study N/A Human: U2OS ATCC Cat#HTB-96; RRID: CVCL_0042 Human: U2OS, FLAG-HaloTag-TRF1 This study N/A Human: U2OS, FLAG-HaloTag-TRF2 This study N/A Human: U2OS, FLAG-HaloTag-TIN2 This study N/A Human: U2OS, FLAG-HaloTag-TPP1 This study N/A Human: U2OS, FLAG-HaloTag-POT1 This study N/A Human: U2OS, FLAG-HaloTag-RAP1 This study N/A Human: U2OS, FLAG-HaloTag-TRF2, RAP1-KO This study N/A Oligonucleotides All oligonucleotides used in this study can be found in Table S2 This study N/A Recombinant DNA pRK2-mNeonGreen-TRF1 plasmid This study N/A pRK2-mNeonGreen-TRF2 plasmid This study N/A pRK2-mNeonGreen-TIN2 plasmid This study N/A pRK2-mNeonGreen-POT1 plasmid This study N/A pFastBacDual plasmid Gibco Cat#10360014 pFastBacDual-FLAG-HaloTag-TRF1 HRD plasmid This study N/A pFastBacDual-FLAG-HaloTag-TRF2 HRD plasmid This study N/A pFastBacDual-FLAG-HaloTag-TIN2 HRD plasmid This study N/A pFastBacDual-FLAG-HaloTag-TPP1 HRD plasmid This study N/A pFastBacDual-FLAG-HaloTag-POT1 HRD plasmid This study N/A pFastBacDual-FLAG-HaloTag-RAP1 HRD plasmid This study N/A pX330-U6-Chimeric_BB-CBh-hSpCas9 plasmid Cong et al. 81 Addgene plasmid #42230 pX330-TRF1 plasmid This study N/A pX330-TRF2 plasmid This study N/A (Continued on next page) Cell Reports 44, 116178, September 23, 2025 17
Recombinant:Article Title: TRF1 and TRF2 form distinct shelterin subcomplexes at telomeres.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse monoclonal anti-TERF2 Novus CatNB100-56506; RRID: AB_839054 Rabbit polyclonal anti-TERF2 Novus Cat#NB110-57130; RRID: AB_844199 Rabbit polyclonal anti-TERF2IP Novus Cat#NB100-292; RRID: AB_10000825 Rabbit polyclonal anti-TPP1 Bethyl Cat#A303-069A; RRID: AB_10890369 Rabbit polyclonal anti-POT1 Proteintech Cat#10581-1-AP; RRID: AB_2284057 Rabbit anti-TERF1 Jan Karlseder N/A Mouse monoclonal anti-FLAG (HRP) Sigma-Aldrich Cat#A8592; RRID: AB_439702 Rabbit polyclonal anti-53BP1 Novus Cat#NB100-304; RRID: AB_10003037 Goat polyclonal anti-rabbit (Cy3) Invitrogen Cat#A10520; RRID: AB_10563288 Goat polyclonal anti-rabbit (HRP) Invitrogen Cat#31460; RRID: AB_228341 Goat polyclonal anti-mouse (HRP) Invitrogen Cat#31439; RRID: AB_228292 Bacterial and virus strains One Shot BL21(DE3) Chemically Competent E. coli Invitrogen Cat#C600003 TOP10 Chemically Competent E. coli Invitrogen Cat#C404006 Chemicals, peptides, and recombinant proteins TEV protease New England Biolabs Cat#P8112 Lipofectamine 2000 Invitrogen Cat#11668500 HaloPROTAC-E AOBIOUS Inc. Cat#AOB13155 Ouabain Sigma-Aldrich Cat#O3125 Thymidine Sigma-Aldrich Cat#T1895 PNA TelC-Alexa488 PNA Bio Cat#F1004 Janelia Fluor JFX650 HaloTag Ligand Grimm et al. 78 ; Promega Cat#HT1070 Janelia Fluor 657 HaloTag Ligand Grimm et al. 79 N/A Janelia Fluor 549 HaloTag Ligand Grimm et al. 80 ; Promega Cat#HT1020 Hoechst 33342 Thermo Scientific Cat#62249 Prolong Diamond Antifade Mountant Invitrogen Cat#P36970 Super Nuclease Sino Biological Cat#SSNP01 ANTI-FLAG M2 Affinity Gel Millipore Cat#A2220 3xFLAG Peptide Sigma-Aldrich Cat#F4799 Ni Sepharose 6 Fast Flow histidine-tagged protein purification resin Cytiva Cat#17531801 Critical commercial assays FxCycle PI/RNase Staining Solution Invitrogen Cat#F10797 SuperSignal West Femto Maximum Sensitivity Substrate Thermo Scientific Cat#34094 4–15% Mini-PROTEAN TGX Stain-Free Protein Gels Bio-Rad Cat#4568083 4–15% Criterion TGX Stain-Free Protein Gel Bio-Rad Cat#5678084 Trans-Blot Turbo RTA Mini 0.2 μm Nitrocellulose Transfer Kit Bio-Rad Cat#1704270 Deposited data All generated plasmids will be deposited at Addgene N/A (Continued on next page) 16 Cell Reports 44, 116178, September 23, 2025 .. REAGENT or RESOURCE SOURCE IDENTIFIER Photobleaching and residence time analysis algorithms This study https://github.com/SchmidtLabMSU/ ShelterinAnalysis Experimental models: Cell lines Human: HeLa EM2-11ht Weidenfeld et al. 50 RRID: CVCL_WN71 Human: HeLa EM2-11ht, FLAG-HaloTag-TRF1 This study N/A Human: HeLa EM2-11ht, FLAG-HaloTag-TRF2 This study N/A Human: HeLa EM2-11ht, FLAG-HaloTag-TIN2 This study N/A Human: HeLa EM2-11ht, FLAG-HaloTag-POT1 This study N/A Human: HeLa EM2-11ht, FLAG-HaloTag-TRF2, RAP1-KO This study N/A Human: HeLa 1.3 Takai et al. 48 RRID: CVCL_B5DA Human: HeLa 1.3, FLAG-HaloTag-TRF2 This study N/A Human: HeLa 1.3, FLAG-HaloTag-TIN2 This study N/A Human: U2OS ATCC Cat#HTB-96; RRID: CVCL_0042 Human: U2OS, FLAG-HaloTag-TRF1 This study N/A Human: U2OS, FLAG-HaloTag-TRF2 This study N/A Human: U2OS, FLAG-HaloTag-TIN2 This study N/A Human: U2OS, FLAG-HaloTag-TPP1 This study N/A Human: U2OS, FLAG-HaloTag-POT1 This study N/A Human: U2OS, FLAG-HaloTag-RAP1 This study N/A Human: U2OS, FLAG-HaloTag-TRF2, RAP1-KO This study N/A Oligonucleotides All oligonucleotides used in this study can be found in Table S2 This study N/A Recombinant DNA pRK2-mNeonGreen-TRF1 plasmid This study N/A pRK2-mNeonGreen-TRF2 plasmid This study N/A pRK2-mNeonGreen-TIN2 plasmid This study N/A pRK2-mNeonGreen-POT1 plasmid This study N/A pFastBacDual plasmid Gibco Cat#10360014 pFastBacDual-FLAG-HaloTag-TRF1 HRD plasmid This study N/A pFastBacDual-FLAG-HaloTag-TRF2 HRD plasmid This study N/A pFastBacDual-FLAG-HaloTag-TIN2 HRD plasmid This study N/A pFastBacDual-FLAG-HaloTag-TPP1 HRD plasmid This study N/A pFastBacDual-FLAG-HaloTag-POT1 HRD plasmid This study N/A pFastBacDual-FLAG-HaloTag-RAP1 HRD plasmid This study N/A pX330-U6-Chimeric_BB-CBh-hSpCas9 plasmid Cong et al. 81 Addgene plasmid #42230 pX330-TRF1 plasmid This study N/A pX330-TRF2 plasmid This study N/A (Continued on next page) Cell Reports 44, 116178, September 23, 2025 17
Protein Purification:Article Title: TRF1 and TRF2 form distinct shelterin subcomplexes at telomeres.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse monoclonal anti-TERF2 Novus CatNB100-56506; RRID: AB_839054 Rabbit polyclonal anti-TERF2 Novus Cat#NB110-57130; RRID: AB_844199 Rabbit polyclonal anti-TERF2IP Novus Cat#NB100-292; RRID: AB_10000825 Rabbit polyclonal anti-TPP1 Bethyl Cat#A303-069A; RRID: AB_10890369 Rabbit polyclonal anti-POT1 Proteintech Cat#10581-1-AP; RRID: AB_2284057 Rabbit anti-TERF1 Jan Karlseder N/A Mouse monoclonal anti-FLAG (HRP) Sigma-Aldrich Cat#A8592; RRID: AB_439702 Rabbit polyclonal anti-53BP1 Novus Cat#NB100-304; RRID: AB_10003037 Goat polyclonal anti-rabbit (Cy3) Invitrogen Cat#A10520; RRID: AB_10563288 Goat polyclonal anti-rabbit (HRP) Invitrogen Cat#31460; RRID: AB_228341 Goat polyclonal anti-mouse (HRP) Invitrogen Cat#31439; RRID: AB_228292 Bacterial and virus strains One Shot BL21(DE3) Chemically Competent E. coli Invitrogen Cat#C600003 TOP10 Chemically Competent E. coli Invitrogen Cat#C404006 Chemicals, peptides, and recombinant proteins TEV protease New England Biolabs Cat#P8112 Lipofectamine 2000 Invitrogen Cat#11668500 HaloPROTAC-E AOBIOUS Inc. Cat#AOB13155 Ouabain Sigma-Aldrich Cat#O3125 Thymidine Sigma-Aldrich Cat#T1895 PNA TelC-Alexa488 PNA Bio Cat#F1004 Janelia Fluor JFX650 HaloTag Ligand Grimm et al. 78 ; Promega Cat#HT1070 Janelia Fluor 657 HaloTag Ligand Grimm et al. 79 N/A Janelia Fluor 549 HaloTag Ligand Grimm et al. 80 ; Promega Cat#HT1020 Hoechst 33342 Thermo Scientific Cat#62249 Prolong Diamond Antifade Mountant Invitrogen Cat#P36970 Super Nuclease Sino Biological Cat#SSNP01 ANTI-FLAG M2 Affinity Gel Millipore Cat#A2220 3xFLAG Peptide Sigma-Aldrich Cat#F4799 Ni Sepharose 6 Fast Flow histidine-tagged protein purification resin Cytiva Cat#17531801 Critical commercial assays FxCycle PI/RNase Staining Solution Invitrogen Cat#F10797 SuperSignal West Femto Maximum Sensitivity Substrate Thermo Scientific Cat#34094 4–15% Mini-PROTEAN TGX Stain-Free Protein Gels Bio-Rad Cat#4568083 4–15% Criterion TGX Stain-Free Protein Gel Bio-Rad Cat#5678084 Trans-Blot Turbo RTA Mini 0.2 μm Nitrocellulose Transfer Kit Bio-Rad Cat#1704270 Deposited data All generated plasmids will be deposited at Addgene N/A (Continued on next page) 16 Cell Reports 44, 116178, September 23, 2025 .. REAGENT or RESOURCE SOURCE IDENTIFIER Photobleaching and residence time analysis algorithms This study https://github.com/SchmidtLabMSU/ ShelterinAnalysis Experimental models: Cell lines Human: HeLa EM2-11ht Weidenfeld et al. 50 RRID: CVCL_WN71 Human: HeLa EM2-11ht, FLAG-HaloTag-TRF1 This study N/A Human: HeLa EM2-11ht, FLAG-HaloTag-TRF2 This study N/A Human: HeLa EM2-11ht, FLAG-HaloTag-TIN2 This study N/A Human: HeLa EM2-11ht, FLAG-HaloTag-POT1 This study N/A Human: HeLa EM2-11ht, FLAG-HaloTag-TRF2, RAP1-KO This study N/A Human: HeLa 1.3 Takai et al. 48 RRID: CVCL_B5DA Human: HeLa 1.3, FLAG-HaloTag-TRF2 This study N/A Human: HeLa 1.3, FLAG-HaloTag-TIN2 This study N/A Human: U2OS ATCC Cat#HTB-96; RRID: CVCL_0042 Human: U2OS, FLAG-HaloTag-TRF1 This study N/A Human: U2OS, FLAG-HaloTag-TRF2 This study N/A Human: U2OS, FLAG-HaloTag-TIN2 This study N/A Human: U2OS, FLAG-HaloTag-TPP1 This study N/A Human: U2OS, FLAG-HaloTag-POT1 This study N/A Human: U2OS, FLAG-HaloTag-RAP1 This study N/A Human: U2OS, FLAG-HaloTag-TRF2, RAP1-KO This study N/A Oligonucleotides All oligonucleotides used in this study can be found in Table S2 This study N/A Recombinant DNA pRK2-mNeonGreen-TRF1 plasmid This study N/A pRK2-mNeonGreen-TRF2 plasmid This study N/A pRK2-mNeonGreen-TIN2 plasmid This study N/A pRK2-mNeonGreen-POT1 plasmid This study N/A pFastBacDual plasmid Gibco Cat#10360014 pFastBacDual-FLAG-HaloTag-TRF1 HRD plasmid This study N/A pFastBacDual-FLAG-HaloTag-TRF2 HRD plasmid This study N/A pFastBacDual-FLAG-HaloTag-TIN2 HRD plasmid This study N/A pFastBacDual-FLAG-HaloTag-TPP1 HRD plasmid This study N/A pFastBacDual-FLAG-HaloTag-POT1 HRD plasmid This study N/A pFastBacDual-FLAG-HaloTag-RAP1 HRD plasmid This study N/A pX330-U6-Chimeric_BB-CBh-hSpCas9 plasmid Cong et al. 81 Addgene plasmid #42230 pX330-TRF1 plasmid This study N/A pX330-TRF2 plasmid This study N/A (Continued on next page) Cell Reports 44, 116178, September 23, 2025 17
Staining:Article Title: TRF1 and TRF2 form distinct shelterin subcomplexes at telomeres.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse monoclonal anti-TERF2 Novus CatNB100-56506; RRID: AB_839054 Rabbit polyclonal anti-TERF2 Novus Cat#NB110-57130; RRID: AB_844199 Rabbit polyclonal anti-TERF2IP Novus Cat#NB100-292; RRID: AB_10000825 Rabbit polyclonal anti-TPP1 Bethyl Cat#A303-069A; RRID: AB_10890369 Rabbit polyclonal anti-POT1 Proteintech Cat#10581-1-AP; RRID: AB_2284057 Rabbit anti-TERF1 Jan Karlseder N/A Mouse monoclonal anti-FLAG (HRP) Sigma-Aldrich Cat#A8592; RRID: AB_439702 Rabbit polyclonal anti-53BP1 Novus Cat#NB100-304; RRID: AB_10003037 Goat polyclonal anti-rabbit (Cy3) Invitrogen Cat#A10520; RRID: AB_10563288 Goat polyclonal anti-rabbit (HRP) Invitrogen Cat#31460; RRID: AB_228341 Goat polyclonal anti-mouse (HRP) Invitrogen Cat#31439; RRID: AB_228292 Bacterial and virus strains One Shot BL21(DE3) Chemically Competent E. coli Invitrogen Cat#C600003 TOP10 Chemically Competent E. coli Invitrogen Cat#C404006 Chemicals, peptides, and recombinant proteins TEV protease New England Biolabs Cat#P8112 Lipofectamine 2000 Invitrogen Cat#11668500 HaloPROTAC-E AOBIOUS Inc. Cat#AOB13155 Ouabain Sigma-Aldrich Cat#O3125 Thymidine Sigma-Aldrich Cat#T1895 PNA TelC-Alexa488 PNA Bio Cat#F1004 Janelia Fluor JFX650 HaloTag Ligand Grimm et al. 78 ; Promega Cat#HT1070 Janelia Fluor 657 HaloTag Ligand Grimm et al. 79 N/A Janelia Fluor 549 HaloTag Ligand Grimm et al. 80 ; Promega Cat#HT1020 Hoechst 33342 Thermo Scientific Cat#62249 Prolong Diamond Antifade Mountant Invitrogen Cat#P36970 Super Nuclease Sino Biological Cat#SSNP01 ANTI-FLAG M2 Affinity Gel Millipore Cat#A2220 3xFLAG Peptide Sigma-Aldrich Cat#F4799 Ni Sepharose 6 Fast Flow histidine-tagged protein purification resin Cytiva Cat#17531801 Critical commercial assays FxCycle PI/RNase Staining Solution Invitrogen Cat#F10797 SuperSignal West Femto Maximum Sensitivity Substrate Thermo Scientific Cat#34094 4–15% Mini-PROTEAN TGX Stain-Free Protein Gels Bio-Rad Cat#4568083 4–15% Criterion TGX Stain-Free Protein Gel Bio-Rad Cat#5678084 Trans-Blot Turbo RTA Mini 0.2 μm Nitrocellulose Transfer Kit Bio-Rad Cat#1704270 Deposited data All generated plasmids will be deposited at Addgene N/A (Continued on next page) 16 Cell Reports 44, 116178, September 23, 2025 .. REAGENT or RESOURCE SOURCE IDENTIFIER Photobleaching and residence time analysis algorithms This study https://github.com/SchmidtLabMSU/ ShelterinAnalysis Experimental models: Cell lines Human: HeLa EM2-11ht Weidenfeld et al. 50 RRID: CVCL_WN71 Human: HeLa EM2-11ht, FLAG-HaloTag-TRF1 This study N/A Human: HeLa EM2-11ht, FLAG-HaloTag-TRF2 This study N/A Human: HeLa EM2-11ht, FLAG-HaloTag-TIN2 This study N/A Human: HeLa EM2-11ht, FLAG-HaloTag-POT1 This study N/A Human: HeLa EM2-11ht, FLAG-HaloTag-TRF2, RAP1-KO This study N/A Human: HeLa 1.3 Takai et al. 48 RRID: CVCL_B5DA Human: HeLa 1.3, FLAG-HaloTag-TRF2 This study N/A Human: HeLa 1.3, FLAG-HaloTag-TIN2 This study N/A Human: U2OS ATCC Cat#HTB-96; RRID: CVCL_0042 Human: U2OS, FLAG-HaloTag-TRF1 This study N/A Human: U2OS, FLAG-HaloTag-TRF2 This study N/A Human: U2OS, FLAG-HaloTag-TIN2 This study N/A Human: U2OS, FLAG-HaloTag-TPP1 This study N/A Human: U2OS, FLAG-HaloTag-POT1 This study N/A Human: U2OS, FLAG-HaloTag-RAP1 This study N/A Human: U2OS, FLAG-HaloTag-TRF2, RAP1-KO This study N/A Oligonucleotides All oligonucleotides used in this study can be found in Table S2 This study N/A Recombinant DNA pRK2-mNeonGreen-TRF1 plasmid This study N/A pRK2-mNeonGreen-TRF2 plasmid This study N/A pRK2-mNeonGreen-TIN2 plasmid This study N/A pRK2-mNeonGreen-POT1 plasmid This study N/A pFastBacDual plasmid Gibco Cat#10360014 pFastBacDual-FLAG-HaloTag-TRF1 HRD plasmid This study N/A pFastBacDual-FLAG-HaloTag-TRF2 HRD plasmid This study N/A pFastBacDual-FLAG-HaloTag-TIN2 HRD plasmid This study N/A pFastBacDual-FLAG-HaloTag-TPP1 HRD plasmid This study N/A pFastBacDual-FLAG-HaloTag-POT1 HRD plasmid This study N/A pFastBacDual-FLAG-HaloTag-RAP1 HRD plasmid This study N/A pX330-U6-Chimeric_BB-CBh-hSpCas9 plasmid Cong et al. 81 Addgene plasmid #42230 pX330-TRF1 plasmid This study N/A pX330-TRF2 plasmid This study N/A (Continued on next page) Cell Reports 44, 116178, September 23, 2025 17
Generated:Article Title: TRF1 and TRF2 form distinct shelterin subcomplexes at telomeres.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse monoclonal anti-TERF2 Novus CatNB100-56506; RRID: AB_839054 Rabbit polyclonal anti-TERF2 Novus Cat#NB110-57130; RRID: AB_844199 Rabbit polyclonal anti-TERF2IP Novus Cat#NB100-292; RRID: AB_10000825 Rabbit polyclonal anti-TPP1 Bethyl Cat#A303-069A; RRID: AB_10890369 Rabbit polyclonal anti-POT1 Proteintech Cat#10581-1-AP; RRID: AB_2284057 Rabbit anti-TERF1 Jan Karlseder N/A Mouse monoclonal anti-FLAG (HRP) Sigma-Aldrich Cat#A8592; RRID: AB_439702 Rabbit polyclonal anti-53BP1 Novus Cat#NB100-304; RRID: AB_10003037 Goat polyclonal anti-rabbit (Cy3) Invitrogen Cat#A10520; RRID: AB_10563288 Goat polyclonal anti-rabbit (HRP) Invitrogen Cat#31460; RRID: AB_228341 Goat polyclonal anti-mouse (HRP) Invitrogen Cat#31439; RRID: AB_228292 Bacterial and virus strains One Shot BL21(DE3) Chemically Competent E. coli Invitrogen Cat#C600003 TOP10 Chemically Competent E. coli Invitrogen Cat#C404006 Chemicals, peptides, and recombinant proteins TEV protease New England Biolabs Cat#P8112 Lipofectamine 2000 Invitrogen Cat#11668500 HaloPROTAC-E AOBIOUS Inc. Cat#AOB13155 Ouabain Sigma-Aldrich Cat#O3125 Thymidine Sigma-Aldrich Cat#T1895 PNA TelC-Alexa488 PNA Bio Cat#F1004 Janelia Fluor JFX650 HaloTag Ligand Grimm et al. 78 ; Promega Cat#HT1070 Janelia Fluor 657 HaloTag Ligand Grimm et al. 79 N/A Janelia Fluor 549 HaloTag Ligand Grimm et al. 80 ; Promega Cat#HT1020 Hoechst 33342 Thermo Scientific Cat#62249 Prolong Diamond Antifade Mountant Invitrogen Cat#P36970 Super Nuclease Sino Biological Cat#SSNP01 ANTI-FLAG M2 Affinity Gel Millipore Cat#A2220 3xFLAG Peptide Sigma-Aldrich Cat#F4799 Ni Sepharose 6 Fast Flow histidine-tagged protein purification resin Cytiva Cat#17531801 Critical commercial assays FxCycle PI/RNase Staining Solution Invitrogen Cat#F10797 SuperSignal West Femto Maximum Sensitivity Substrate Thermo Scientific Cat#34094 4–15% Mini-PROTEAN TGX Stain-Free Protein Gels Bio-Rad Cat#4568083 4–15% Criterion TGX Stain-Free Protein Gel Bio-Rad Cat#5678084 Trans-Blot Turbo RTA Mini 0.2 μm Nitrocellulose Transfer Kit Bio-Rad Cat#1704270 Deposited data All generated plasmids will be deposited at Addgene N/A (Continued on next page) 16 Cell Reports 44, 116178, September 23, 2025 .. REAGENT or RESOURCE SOURCE IDENTIFIER Photobleaching and residence time analysis algorithms This study https://github.com/SchmidtLabMSU/ ShelterinAnalysis Experimental models: Cell lines Human: HeLa EM2-11ht Weidenfeld et al. 50 RRID: CVCL_WN71 Human: HeLa EM2-11ht, FLAG-HaloTag-TRF1 This study N/A Human: HeLa EM2-11ht, FLAG-HaloTag-TRF2 This study N/A Human: HeLa EM2-11ht, FLAG-HaloTag-TIN2 This study N/A Human: HeLa EM2-11ht, FLAG-HaloTag-POT1 This study N/A Human: HeLa EM2-11ht, FLAG-HaloTag-TRF2, RAP1-KO This study N/A Human: HeLa 1.3 Takai et al. 48 RRID: CVCL_B5DA Human: HeLa 1.3, FLAG-HaloTag-TRF2 This study N/A Human: HeLa 1.3, FLAG-HaloTag-TIN2 This study N/A Human: U2OS ATCC Cat#HTB-96; RRID: CVCL_0042 Human: U2OS, FLAG-HaloTag-TRF1 This study N/A Human: U2OS, FLAG-HaloTag-TRF2 This study N/A Human: U2OS, FLAG-HaloTag-TIN2 This study N/A Human: U2OS, FLAG-HaloTag-TPP1 This study N/A Human: U2OS, FLAG-HaloTag-POT1 This study N/A Human: U2OS, FLAG-HaloTag-RAP1 This study N/A Human: U2OS, FLAG-HaloTag-TRF2, RAP1-KO This study N/A Oligonucleotides All oligonucleotides used in this study can be found in Table S2 This study N/A Recombinant DNA pRK2-mNeonGreen-TRF1 plasmid This study N/A pRK2-mNeonGreen-TRF2 plasmid This study N/A pRK2-mNeonGreen-TIN2 plasmid This study N/A pRK2-mNeonGreen-POT1 plasmid This study N/A pFastBacDual plasmid Gibco Cat#10360014 pFastBacDual-FLAG-HaloTag-TRF1 HRD plasmid This study N/A pFastBacDual-FLAG-HaloTag-TRF2 HRD plasmid This study N/A pFastBacDual-FLAG-HaloTag-TIN2 HRD plasmid This study N/A pFastBacDual-FLAG-HaloTag-TPP1 HRD plasmid This study N/A pFastBacDual-FLAG-HaloTag-POT1 HRD plasmid This study N/A pFastBacDual-FLAG-HaloTag-RAP1 HRD plasmid This study N/A pX330-U6-Chimeric_BB-CBh-hSpCas9 plasmid Cong et al. 81 Addgene plasmid #42230 pX330-TRF1 plasmid This study N/A pX330-TRF2 plasmid This study N/A (Continued on next page) Cell Reports 44, 116178, September 23, 2025 17
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