Recombinant:Article Title: SMUG1 Promotes Telomere Maintenance through Telomerase RNA Processing.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER mouse IL-6 recombinant protein eBioscience Cat# 14-8061-80 mouse stem cell factor (SCF) recombinant protein eBioscience Cat# 14-8341-63 Mouse Methylcellulose Complete Media R&D systems Cat# HSC007 Colcemid Life Technologies Cat# 15210040 Blocking reagent Sigma-Aldrich-Aldrich Cat# 11096176001 Vectashield mounting medium with DAPI Vector labs Cat# H-1200 Pierce BCA Protein Assay Kit Life Technologies Cat# 23227 DNeasy Blood & Tissue Kit QIAGEN Cat# 69504 APE1 New England Biolabs Cat# M0282S Thymidine Sigma-Aldrich-Aldrich Cat# T1895 Imidazole buffer Solution Sigma-Aldrich-Aldrich Cat# 68268 Isopropyl b-D-1-thiogalactopyranoside Sigma-Aldrich-Aldrich Cat# I5502 Ni-NTA agarose resin QIAGEN Cat# 30210 Econo-Column Chromatography Columns Bio-Rad Cat# 7372522 Bio-Scale Mini Macro-Prep High S cartridge Bio-Rad Cat# 7324134 Ultrafree-MC Centrifugal Filter Millipore Cat# UFC30DV0S HIS-Select Nickel Magnetic Agarose beads Sigma-Aldrich-Aldrich Cat# H9914 SMUG1 wt recombinant protein Lab made N/A SMUG1 H239L recombinant protein Lab made N/A Critical Commercial Assays SENSE Total RNA-Seq Library Prep Kit Lexogen Cat# SKU: 009.24 Click-iT Nascent RNA Capture Kit, for gene expression analysis Life Technologies Cat# C10365 DIG Oligonucleotide 30 End Labeling Kit, 2nd generation Sigma-Aldrich-Aldrich Cat# 03353575910 TeloTAGGG Telomere Length Assay Sigma-Aldrich-Aldrich Cat# 12209136001 TruSeq Nano DNA Low Throughput Library Prep Kit Illumina Cat# 20015964 MiSeq Reagent Kit v2 (500-cycles) Illumina Cat# MS-102-2003 Duolink In Situ Red Starter Kit Mouse/Rabbit Sigma-Aldrich-Aldrich Cat# DUO92101 TRAPeze Telomerase Detection Kit Millipore Cat# S7710 MEGAscript T7 Transcription Kit Life Technologies Cat# AM1333 Deposited Data RNA-seq data This study GEO: GSE116580 30 RACE seq data This study GEO: GSE116580 Experimental Models: Cell Lines Mouse: Mouse Embryonic fibroblasts Smug1+/+ Lab made Alsøe et al., 2017 Mouse: Mouse Embryonic fibroblasts Smug1 / Lab made Alsøe et al., 2017 Mouse: Mouse Embryonic fibroblasts Smug1 / _SMUG1 Lab made N/A Mouse: Mouse Embryonic fibroblasts Smug1 / _H241L Lab made N/A Mouse: Mouse Embryonic fibroblasts Smug1 / _E31R/E35R Lab made N/A Human: HAP1 SMUG1-WT (control wild type cell line) Horizon Cat# HZGHC003300c009 Human: HAP1 SMUG1-KO Horizon Cat# HZGHC003300c009 Human: HAP1 SMUG1-KO_SMUG1 Lab made N/A Human: HAP1 SMUG1-KO_SMUG1 H239L (NABm) Lab made N/A Human: HAP1 SMUG1-KO_E29R/E33R/E231R (DBm) Lab made N/A (Continued on next page) Cell Reports 28, 1690–1702.e1–e10, August 13, 2019 e3 .. REAGENT or RESOURCE SOURCE IDENTIFIER Human: HeLa ATCC ATCC CCL-2 Experimental Models: Organisms/Strains C56BL/6 Smug1tm1Hln (Smug1 / ) Lab made Alsøe et al., 2017 Oligonucleotides See Table S1 for oligonucleotide information N/A Recombinant DNA Plasmid: pHH25-Smug1 Cloned and stored in lab N/A Plasmid: pHH25-Smug1 H241L Cloned and stored in lab N/A Plasmid: pHH25-Smug1 E31R/E35R Cloned and stored in lab N/A Plasmid: pHH25-SMUG1 Cloned and stored in lab N/A Plasmid: pHH25-SMUG1 H239L (NABm) Cloned and stored in lab N/A Plasmid: pHH25-SMUG1 E29R/E31R/E231R (DBm) Cloned and stored in lab N/A Plasmid: pCDNA-3xHA-hTERT Addgene Cat# 51631; RRID:Addgene_51637 Plasmid: pBS U3-hTR-500 Addgene Cat# 28170; RRID:Addgene_28170 Plasmid: pETM-11-hSMUG1 wt Cloned and stored in lab N/A Plasmid: pETM-11-hSMUG1 H239L Cloned and stored in lab N/A Software and Algorithms Multi Gauge V3.1 software Fujifilm N/A STAR aligner Dobin et al., 2013 https://github.com/alexdobin/STAR; RRID:SCR_015899 htseq-count Anders et al., 2015 https://github.com/simon-anders/htseq; RRID:SCR_011867 Voom Law et al., 2014 http://bioinf.wehi.edu.au/voom/ Limma Ritchie et al., 2015 https://bioconductor.org/packages/ release/bioc/html/limma.html; RRID:SCR_010943 ImageJ https://imagej.net/Welcome; RRID:SCR_003070 bowtie2 Langmead and Salzberg, 2012 http://bowtie-bio.sourceforge.net/bowtie2/ index.shtml Bedtools Quinlan and Hall, 2010 https://github.com/arq5x/bedtools2; RRID:SCR_006646 TeloTool Göhring et al., 2014 https://github.com/jagoehring/TeloTool Zeiss Zen blue and black Zeiss https://www.zeiss.com/microscopy/en_us/ products/microscope-software/zen.html; RRID:SCR_013672 GraphPad Prism GraphPad https://www.graphpad.com/; RRID:SCR_002798 EZLogic Integration Bio-Rad http://www.bio-rad.com
Blocking Assay:Article Title: SMUG1 Promotes Telomere Maintenance through Telomerase RNA Processing.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER mouse IL-6 recombinant protein eBioscience Cat# 14-8061-80 mouse stem cell factor (SCF) recombinant protein eBioscience Cat# 14-8341-63 Mouse Methylcellulose Complete Media R&D systems Cat# HSC007 Colcemid Life Technologies Cat# 15210040 Blocking reagent Sigma-Aldrich-Aldrich Cat# 11096176001 Vectashield mounting medium with DAPI Vector labs Cat# H-1200 Pierce BCA Protein Assay Kit Life Technologies Cat# 23227 DNeasy Blood & Tissue Kit QIAGEN Cat# 69504 APE1 New England Biolabs Cat# M0282S Thymidine Sigma-Aldrich-Aldrich Cat# T1895 Imidazole buffer Solution Sigma-Aldrich-Aldrich Cat# 68268 Isopropyl b-D-1-thiogalactopyranoside Sigma-Aldrich-Aldrich Cat# I5502 Ni-NTA agarose resin QIAGEN Cat# 30210 Econo-Column Chromatography Columns Bio-Rad Cat# 7372522 Bio-Scale Mini Macro-Prep High S cartridge Bio-Rad Cat# 7324134 Ultrafree-MC Centrifugal Filter Millipore Cat# UFC30DV0S HIS-Select Nickel Magnetic Agarose beads Sigma-Aldrich-Aldrich Cat# H9914 SMUG1 wt recombinant protein Lab made N/A SMUG1 H239L recombinant protein Lab made N/A Critical Commercial Assays SENSE Total RNA-Seq Library Prep Kit Lexogen Cat# SKU: 009.24 Click-iT Nascent RNA Capture Kit, for gene expression analysis Life Technologies Cat# C10365 DIG Oligonucleotide 30 End Labeling Kit, 2nd generation Sigma-Aldrich-Aldrich Cat# 03353575910 TeloTAGGG Telomere Length Assay Sigma-Aldrich-Aldrich Cat# 12209136001 TruSeq Nano DNA Low Throughput Library Prep Kit Illumina Cat# 20015964 MiSeq Reagent Kit v2 (500-cycles) Illumina Cat# MS-102-2003 Duolink In Situ Red Starter Kit Mouse/Rabbit Sigma-Aldrich-Aldrich Cat# DUO92101 TRAPeze Telomerase Detection Kit Millipore Cat# S7710 MEGAscript T7 Transcription Kit Life Technologies Cat# AM1333 Deposited Data RNA-seq data This study GEO: GSE116580 30 RACE seq data This study GEO: GSE116580 Experimental Models: Cell Lines Mouse: Mouse Embryonic fibroblasts Smug1+/+ Lab made Alsøe et al., 2017 Mouse: Mouse Embryonic fibroblasts Smug1 / Lab made Alsøe et al., 2017 Mouse: Mouse Embryonic fibroblasts Smug1 / _SMUG1 Lab made N/A Mouse: Mouse Embryonic fibroblasts Smug1 / _H241L Lab made N/A Mouse: Mouse Embryonic fibroblasts Smug1 / _E31R/E35R Lab made N/A Human: HAP1 SMUG1-WT (control wild type cell line) Horizon Cat# HZGHC003300c009 Human: HAP1 SMUG1-KO Horizon Cat# HZGHC003300c009 Human: HAP1 SMUG1-KO_SMUG1 Lab made N/A Human: HAP1 SMUG1-KO_SMUG1 H239L (NABm) Lab made N/A Human: HAP1 SMUG1-KO_E29R/E33R/E231R (DBm) Lab made N/A (Continued on next page) Cell Reports 28, 1690–1702.e1–e10, August 13, 2019 e3 .. REAGENT or RESOURCE SOURCE IDENTIFIER Human: HeLa ATCC ATCC CCL-2 Experimental Models: Organisms/Strains C56BL/6 Smug1tm1Hln (Smug1 / ) Lab made Alsøe et al., 2017 Oligonucleotides See Table S1 for oligonucleotide information N/A Recombinant DNA Plasmid: pHH25-Smug1 Cloned and stored in lab N/A Plasmid: pHH25-Smug1 H241L Cloned and stored in lab N/A Plasmid: pHH25-Smug1 E31R/E35R Cloned and stored in lab N/A Plasmid: pHH25-SMUG1 Cloned and stored in lab N/A Plasmid: pHH25-SMUG1 H239L (NABm) Cloned and stored in lab N/A Plasmid: pHH25-SMUG1 E29R/E31R/E231R (DBm) Cloned and stored in lab N/A Plasmid: pCDNA-3xHA-hTERT Addgene Cat# 51631; RRID:Addgene_51637 Plasmid: pBS U3-hTR-500 Addgene Cat# 28170; RRID:Addgene_28170 Plasmid: pETM-11-hSMUG1 wt Cloned and stored in lab N/A Plasmid: pETM-11-hSMUG1 H239L Cloned and stored in lab N/A Software and Algorithms Multi Gauge V3.1 software Fujifilm N/A STAR aligner Dobin et al., 2013 https://github.com/alexdobin/STAR; RRID:SCR_015899 htseq-count Anders et al., 2015 https://github.com/simon-anders/htseq; RRID:SCR_011867 Voom Law et al., 2014 http://bioinf.wehi.edu.au/voom/ Limma Ritchie et al., 2015 https://bioconductor.org/packages/ release/bioc/html/limma.html; RRID:SCR_010943 ImageJ https://imagej.net/Welcome; RRID:SCR_003070 bowtie2 Langmead and Salzberg, 2012 http://bowtie-bio.sourceforge.net/bowtie2/ index.shtml Bedtools Quinlan and Hall, 2010 https://github.com/arq5x/bedtools2; RRID:SCR_006646 TeloTool Göhring et al., 2014 https://github.com/jagoehring/TeloTool Zeiss Zen blue and black Zeiss https://www.zeiss.com/microscopy/en_us/ products/microscope-software/zen.html; RRID:SCR_013672 GraphPad Prism GraphPad https://www.graphpad.com/; RRID:SCR_002798 EZLogic Integration Bio-Rad http://www.bio-rad.com
Bicinchoninic Acid Protein Assay:Article Title: SMUG1 Promotes Telomere Maintenance through Telomerase RNA Processing.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER mouse IL-6 recombinant protein eBioscience Cat# 14-8061-80 mouse stem cell factor (SCF) recombinant protein eBioscience Cat# 14-8341-63 Mouse Methylcellulose Complete Media R&D systems Cat# HSC007 Colcemid Life Technologies Cat# 15210040 Blocking reagent Sigma-Aldrich-Aldrich Cat# 11096176001 Vectashield mounting medium with DAPI Vector labs Cat# H-1200 Pierce BCA Protein Assay Kit Life Technologies Cat# 23227 DNeasy Blood & Tissue Kit QIAGEN Cat# 69504 APE1 New England Biolabs Cat# M0282S Thymidine Sigma-Aldrich-Aldrich Cat# T1895 Imidazole buffer Solution Sigma-Aldrich-Aldrich Cat# 68268 Isopropyl b-D-1-thiogalactopyranoside Sigma-Aldrich-Aldrich Cat# I5502 Ni-NTA agarose resin QIAGEN Cat# 30210 Econo-Column Chromatography Columns Bio-Rad Cat# 7372522 Bio-Scale Mini Macro-Prep High S cartridge Bio-Rad Cat# 7324134 Ultrafree-MC Centrifugal Filter Millipore Cat# UFC30DV0S HIS-Select Nickel Magnetic Agarose beads Sigma-Aldrich-Aldrich Cat# H9914 SMUG1 wt recombinant protein Lab made N/A SMUG1 H239L recombinant protein Lab made N/A Critical Commercial Assays SENSE Total RNA-Seq Library Prep Kit Lexogen Cat# SKU: 009.24 Click-iT Nascent RNA Capture Kit, for gene expression analysis Life Technologies Cat# C10365 DIG Oligonucleotide 30 End Labeling Kit, 2nd generation Sigma-Aldrich-Aldrich Cat# 03353575910 TeloTAGGG Telomere Length Assay Sigma-Aldrich-Aldrich Cat# 12209136001 TruSeq Nano DNA Low Throughput Library Prep Kit Illumina Cat# 20015964 MiSeq Reagent Kit v2 (500-cycles) Illumina Cat# MS-102-2003 Duolink In Situ Red Starter Kit Mouse/Rabbit Sigma-Aldrich-Aldrich Cat# DUO92101 TRAPeze Telomerase Detection Kit Millipore Cat# S7710 MEGAscript T7 Transcription Kit Life Technologies Cat# AM1333 Deposited Data RNA-seq data This study GEO: GSE116580 30 RACE seq data This study GEO: GSE116580 Experimental Models: Cell Lines Mouse: Mouse Embryonic fibroblasts Smug1+/+ Lab made Alsøe et al., 2017 Mouse: Mouse Embryonic fibroblasts Smug1 / Lab made Alsøe et al., 2017 Mouse: Mouse Embryonic fibroblasts Smug1 / _SMUG1 Lab made N/A Mouse: Mouse Embryonic fibroblasts Smug1 / _H241L Lab made N/A Mouse: Mouse Embryonic fibroblasts Smug1 / _E31R/E35R Lab made N/A Human: HAP1 SMUG1-WT (control wild type cell line) Horizon Cat# HZGHC003300c009 Human: HAP1 SMUG1-KO Horizon Cat# HZGHC003300c009 Human: HAP1 SMUG1-KO_SMUG1 Lab made N/A Human: HAP1 SMUG1-KO_SMUG1 H239L (NABm) Lab made N/A Human: HAP1 SMUG1-KO_E29R/E33R/E231R (DBm) Lab made N/A (Continued on next page) Cell Reports 28, 1690–1702.e1–e10, August 13, 2019 e3 .. REAGENT or RESOURCE SOURCE IDENTIFIER Human: HeLa ATCC ATCC CCL-2 Experimental Models: Organisms/Strains C56BL/6 Smug1tm1Hln (Smug1 / ) Lab made Alsøe et al., 2017 Oligonucleotides See Table S1 for oligonucleotide information N/A Recombinant DNA Plasmid: pHH25-Smug1 Cloned and stored in lab N/A Plasmid: pHH25-Smug1 H241L Cloned and stored in lab N/A Plasmid: pHH25-Smug1 E31R/E35R Cloned and stored in lab N/A Plasmid: pHH25-SMUG1 Cloned and stored in lab N/A Plasmid: pHH25-SMUG1 H239L (NABm) Cloned and stored in lab N/A Plasmid: pHH25-SMUG1 E29R/E31R/E231R (DBm) Cloned and stored in lab N/A Plasmid: pCDNA-3xHA-hTERT Addgene Cat# 51631; RRID:Addgene_51637 Plasmid: pBS U3-hTR-500 Addgene Cat# 28170; RRID:Addgene_28170 Plasmid: pETM-11-hSMUG1 wt Cloned and stored in lab N/A Plasmid: pETM-11-hSMUG1 H239L Cloned and stored in lab N/A Software and Algorithms Multi Gauge V3.1 software Fujifilm N/A STAR aligner Dobin et al., 2013 https://github.com/alexdobin/STAR; RRID:SCR_015899 htseq-count Anders et al., 2015 https://github.com/simon-anders/htseq; RRID:SCR_011867 Voom Law et al., 2014 http://bioinf.wehi.edu.au/voom/ Limma Ritchie et al., 2015 https://bioconductor.org/packages/ release/bioc/html/limma.html; RRID:SCR_010943 ImageJ https://imagej.net/Welcome; RRID:SCR_003070 bowtie2 Langmead and Salzberg, 2012 http://bowtie-bio.sourceforge.net/bowtie2/ index.shtml Bedtools Quinlan and Hall, 2010 https://github.com/arq5x/bedtools2; RRID:SCR_006646 TeloTool Göhring et al., 2014 https://github.com/jagoehring/TeloTool Zeiss Zen blue and black Zeiss https://www.zeiss.com/microscopy/en_us/ products/microscope-software/zen.html; RRID:SCR_013672 GraphPad Prism GraphPad https://www.graphpad.com/; RRID:SCR_002798 EZLogic Integration Bio-Rad http://www.bio-rad.com
Chromatography:Article Title: SMUG1 Promotes Telomere Maintenance through Telomerase RNA Processing.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER mouse IL-6 recombinant protein eBioscience Cat# 14-8061-80 mouse stem cell factor (SCF) recombinant protein eBioscience Cat# 14-8341-63 Mouse Methylcellulose Complete Media R&D systems Cat# HSC007 Colcemid Life Technologies Cat# 15210040 Blocking reagent Sigma-Aldrich-Aldrich Cat# 11096176001 Vectashield mounting medium with DAPI Vector labs Cat# H-1200 Pierce BCA Protein Assay Kit Life Technologies Cat# 23227 DNeasy Blood & Tissue Kit QIAGEN Cat# 69504 APE1 New England Biolabs Cat# M0282S Thymidine Sigma-Aldrich-Aldrich Cat# T1895 Imidazole buffer Solution Sigma-Aldrich-Aldrich Cat# 68268 Isopropyl b-D-1-thiogalactopyranoside Sigma-Aldrich-Aldrich Cat# I5502 Ni-NTA agarose resin QIAGEN Cat# 30210 Econo-Column Chromatography Columns Bio-Rad Cat# 7372522 Bio-Scale Mini Macro-Prep High S cartridge Bio-Rad Cat# 7324134 Ultrafree-MC Centrifugal Filter Millipore Cat# UFC30DV0S HIS-Select Nickel Magnetic Agarose beads Sigma-Aldrich-Aldrich Cat# H9914 SMUG1 wt recombinant protein Lab made N/A SMUG1 H239L recombinant protein Lab made N/A Critical Commercial Assays SENSE Total RNA-Seq Library Prep Kit Lexogen Cat# SKU: 009.24 Click-iT Nascent RNA Capture Kit, for gene expression analysis Life Technologies Cat# C10365 DIG Oligonucleotide 30 End Labeling Kit, 2nd generation Sigma-Aldrich-Aldrich Cat# 03353575910 TeloTAGGG Telomere Length Assay Sigma-Aldrich-Aldrich Cat# 12209136001 TruSeq Nano DNA Low Throughput Library Prep Kit Illumina Cat# 20015964 MiSeq Reagent Kit v2 (500-cycles) Illumina Cat# MS-102-2003 Duolink In Situ Red Starter Kit Mouse/Rabbit Sigma-Aldrich-Aldrich Cat# DUO92101 TRAPeze Telomerase Detection Kit Millipore Cat# S7710 MEGAscript T7 Transcription Kit Life Technologies Cat# AM1333 Deposited Data RNA-seq data This study GEO: GSE116580 30 RACE seq data This study GEO: GSE116580 Experimental Models: Cell Lines Mouse: Mouse Embryonic fibroblasts Smug1+/+ Lab made Alsøe et al., 2017 Mouse: Mouse Embryonic fibroblasts Smug1 / Lab made Alsøe et al., 2017 Mouse: Mouse Embryonic fibroblasts Smug1 / _SMUG1 Lab made N/A Mouse: Mouse Embryonic fibroblasts Smug1 / _H241L Lab made N/A Mouse: Mouse Embryonic fibroblasts Smug1 / _E31R/E35R Lab made N/A Human: HAP1 SMUG1-WT (control wild type cell line) Horizon Cat# HZGHC003300c009 Human: HAP1 SMUG1-KO Horizon Cat# HZGHC003300c009 Human: HAP1 SMUG1-KO_SMUG1 Lab made N/A Human: HAP1 SMUG1-KO_SMUG1 H239L (NABm) Lab made N/A Human: HAP1 SMUG1-KO_E29R/E33R/E231R (DBm) Lab made N/A (Continued on next page) Cell Reports 28, 1690–1702.e1–e10, August 13, 2019 e3 .. REAGENT or RESOURCE SOURCE IDENTIFIER Human: HeLa ATCC ATCC CCL-2 Experimental Models: Organisms/Strains C56BL/6 Smug1tm1Hln (Smug1 / ) Lab made Alsøe et al., 2017 Oligonucleotides See Table S1 for oligonucleotide information N/A Recombinant DNA Plasmid: pHH25-Smug1 Cloned and stored in lab N/A Plasmid: pHH25-Smug1 H241L Cloned and stored in lab N/A Plasmid: pHH25-Smug1 E31R/E35R Cloned and stored in lab N/A Plasmid: pHH25-SMUG1 Cloned and stored in lab N/A Plasmid: pHH25-SMUG1 H239L (NABm) Cloned and stored in lab N/A Plasmid: pHH25-SMUG1 E29R/E31R/E231R (DBm) Cloned and stored in lab N/A Plasmid: pCDNA-3xHA-hTERT Addgene Cat# 51631; RRID:Addgene_51637 Plasmid: pBS U3-hTR-500 Addgene Cat# 28170; RRID:Addgene_28170 Plasmid: pETM-11-hSMUG1 wt Cloned and stored in lab N/A Plasmid: pETM-11-hSMUG1 H239L Cloned and stored in lab N/A Software and Algorithms Multi Gauge V3.1 software Fujifilm N/A STAR aligner Dobin et al., 2013 https://github.com/alexdobin/STAR; RRID:SCR_015899 htseq-count Anders et al., 2015 https://github.com/simon-anders/htseq; RRID:SCR_011867 Voom Law et al., 2014 http://bioinf.wehi.edu.au/voom/ Limma Ritchie et al., 2015 https://bioconductor.org/packages/ release/bioc/html/limma.html; RRID:SCR_010943 ImageJ https://imagej.net/Welcome; RRID:SCR_003070 bowtie2 Langmead and Salzberg, 2012 http://bowtie-bio.sourceforge.net/bowtie2/ index.shtml Bedtools Quinlan and Hall, 2010 https://github.com/arq5x/bedtools2; RRID:SCR_006646 TeloTool Göhring et al., 2014 https://github.com/jagoehring/TeloTool Zeiss Zen blue and black Zeiss https://www.zeiss.com/microscopy/en_us/ products/microscope-software/zen.html; RRID:SCR_013672 GraphPad Prism GraphPad https://www.graphpad.com/; RRID:SCR_002798 EZLogic Integration Bio-Rad http://www.bio-rad.com
RNA sequencing:Article Title: SMUG1 Promotes Telomere Maintenance through Telomerase RNA Processing.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER mouse IL-6 recombinant protein eBioscience Cat# 14-8061-80 mouse stem cell factor (SCF) recombinant protein eBioscience Cat# 14-8341-63 Mouse Methylcellulose Complete Media R&D systems Cat# HSC007 Colcemid Life Technologies Cat# 15210040 Blocking reagent Sigma-Aldrich-Aldrich Cat# 11096176001 Vectashield mounting medium with DAPI Vector labs Cat# H-1200 Pierce BCA Protein Assay Kit Life Technologies Cat# 23227 DNeasy Blood & Tissue Kit QIAGEN Cat# 69504 APE1 New England Biolabs Cat# M0282S Thymidine Sigma-Aldrich-Aldrich Cat# T1895 Imidazole buffer Solution Sigma-Aldrich-Aldrich Cat# 68268 Isopropyl b-D-1-thiogalactopyranoside Sigma-Aldrich-Aldrich Cat# I5502 Ni-NTA agarose resin QIAGEN Cat# 30210 Econo-Column Chromatography Columns Bio-Rad Cat# 7372522 Bio-Scale Mini Macro-Prep High S cartridge Bio-Rad Cat# 7324134 Ultrafree-MC Centrifugal Filter Millipore Cat# UFC30DV0S HIS-Select Nickel Magnetic Agarose beads Sigma-Aldrich-Aldrich Cat# H9914 SMUG1 wt recombinant protein Lab made N/A SMUG1 H239L recombinant protein Lab made N/A Critical Commercial Assays SENSE Total RNA-Seq Library Prep Kit Lexogen Cat# SKU: 009.24 Click-iT Nascent RNA Capture Kit, for gene expression analysis Life Technologies Cat# C10365 DIG Oligonucleotide 30 End Labeling Kit, 2nd generation Sigma-Aldrich-Aldrich Cat# 03353575910 TeloTAGGG Telomere Length Assay Sigma-Aldrich-Aldrich Cat# 12209136001 TruSeq Nano DNA Low Throughput Library Prep Kit Illumina Cat# 20015964 MiSeq Reagent Kit v2 (500-cycles) Illumina Cat# MS-102-2003 Duolink In Situ Red Starter Kit Mouse/Rabbit Sigma-Aldrich-Aldrich Cat# DUO92101 TRAPeze Telomerase Detection Kit Millipore Cat# S7710 MEGAscript T7 Transcription Kit Life Technologies Cat# AM1333 Deposited Data RNA-seq data This study GEO: GSE116580 30 RACE seq data This study GEO: GSE116580 Experimental Models: Cell Lines Mouse: Mouse Embryonic fibroblasts Smug1+/+ Lab made Alsøe et al., 2017 Mouse: Mouse Embryonic fibroblasts Smug1 / Lab made Alsøe et al., 2017 Mouse: Mouse Embryonic fibroblasts Smug1 / _SMUG1 Lab made N/A Mouse: Mouse Embryonic fibroblasts Smug1 / _H241L Lab made N/A Mouse: Mouse Embryonic fibroblasts Smug1 / _E31R/E35R Lab made N/A Human: HAP1 SMUG1-WT (control wild type cell line) Horizon Cat# HZGHC003300c009 Human: HAP1 SMUG1-KO Horizon Cat# HZGHC003300c009 Human: HAP1 SMUG1-KO_SMUG1 Lab made N/A Human: HAP1 SMUG1-KO_SMUG1 H239L (NABm) Lab made N/A Human: HAP1 SMUG1-KO_E29R/E33R/E231R (DBm) Lab made N/A (Continued on next page) Cell Reports 28, 1690–1702.e1–e10, August 13, 2019 e3 .. REAGENT or RESOURCE SOURCE IDENTIFIER Human: HeLa ATCC ATCC CCL-2 Experimental Models: Organisms/Strains C56BL/6 Smug1tm1Hln (Smug1 / ) Lab made Alsøe et al., 2017 Oligonucleotides See Table S1 for oligonucleotide information N/A Recombinant DNA Plasmid: pHH25-Smug1 Cloned and stored in lab N/A Plasmid: pHH25-Smug1 H241L Cloned and stored in lab N/A Plasmid: pHH25-Smug1 E31R/E35R Cloned and stored in lab N/A Plasmid: pHH25-SMUG1 Cloned and stored in lab N/A Plasmid: pHH25-SMUG1 H239L (NABm) Cloned and stored in lab N/A Plasmid: pHH25-SMUG1 E29R/E31R/E231R (DBm) Cloned and stored in lab N/A Plasmid: pCDNA-3xHA-hTERT Addgene Cat# 51631; RRID:Addgene_51637 Plasmid: pBS U3-hTR-500 Addgene Cat# 28170; RRID:Addgene_28170 Plasmid: pETM-11-hSMUG1 wt Cloned and stored in lab N/A Plasmid: pETM-11-hSMUG1 H239L Cloned and stored in lab N/A Software and Algorithms Multi Gauge V3.1 software Fujifilm N/A STAR aligner Dobin et al., 2013 https://github.com/alexdobin/STAR; RRID:SCR_015899 htseq-count Anders et al., 2015 https://github.com/simon-anders/htseq; RRID:SCR_011867 Voom Law et al., 2014 http://bioinf.wehi.edu.au/voom/ Limma Ritchie et al., 2015 https://bioconductor.org/packages/ release/bioc/html/limma.html; RRID:SCR_010943 ImageJ https://imagej.net/Welcome; RRID:SCR_003070 bowtie2 Langmead and Salzberg, 2012 http://bowtie-bio.sourceforge.net/bowtie2/ index.shtml Bedtools Quinlan and Hall, 2010 https://github.com/arq5x/bedtools2; RRID:SCR_006646 TeloTool Göhring et al., 2014 https://github.com/jagoehring/TeloTool Zeiss Zen blue and black Zeiss https://www.zeiss.com/microscopy/en_us/ products/microscope-software/zen.html; RRID:SCR_013672 GraphPad Prism GraphPad https://www.graphpad.com/; RRID:SCR_002798 EZLogic Integration Bio-Rad http://www.bio-rad.com
Gene Expression:Article Title: SMUG1 Promotes Telomere Maintenance through Telomerase RNA Processing.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER mouse IL-6 recombinant protein eBioscience Cat# 14-8061-80 mouse stem cell factor (SCF) recombinant protein eBioscience Cat# 14-8341-63 Mouse Methylcellulose Complete Media R&D systems Cat# HSC007 Colcemid Life Technologies Cat# 15210040 Blocking reagent Sigma-Aldrich-Aldrich Cat# 11096176001 Vectashield mounting medium with DAPI Vector labs Cat# H-1200 Pierce BCA Protein Assay Kit Life Technologies Cat# 23227 DNeasy Blood & Tissue Kit QIAGEN Cat# 69504 APE1 New England Biolabs Cat# M0282S Thymidine Sigma-Aldrich-Aldrich Cat# T1895 Imidazole buffer Solution Sigma-Aldrich-Aldrich Cat# 68268 Isopropyl b-D-1-thiogalactopyranoside Sigma-Aldrich-Aldrich Cat# I5502 Ni-NTA agarose resin QIAGEN Cat# 30210 Econo-Column Chromatography Columns Bio-Rad Cat# 7372522 Bio-Scale Mini Macro-Prep High S cartridge Bio-Rad Cat# 7324134 Ultrafree-MC Centrifugal Filter Millipore Cat# UFC30DV0S HIS-Select Nickel Magnetic Agarose beads Sigma-Aldrich-Aldrich Cat# H9914 SMUG1 wt recombinant protein Lab made N/A SMUG1 H239L recombinant protein Lab made N/A Critical Commercial Assays SENSE Total RNA-Seq Library Prep Kit Lexogen Cat# SKU: 009.24 Click-iT Nascent RNA Capture Kit, for gene expression analysis Life Technologies Cat# C10365 DIG Oligonucleotide 30 End Labeling Kit, 2nd generation Sigma-Aldrich-Aldrich Cat# 03353575910 TeloTAGGG Telomere Length Assay Sigma-Aldrich-Aldrich Cat# 12209136001 TruSeq Nano DNA Low Throughput Library Prep Kit Illumina Cat# 20015964 MiSeq Reagent Kit v2 (500-cycles) Illumina Cat# MS-102-2003 Duolink In Situ Red Starter Kit Mouse/Rabbit Sigma-Aldrich-Aldrich Cat# DUO92101 TRAPeze Telomerase Detection Kit Millipore Cat# S7710 MEGAscript T7 Transcription Kit Life Technologies Cat# AM1333 Deposited Data RNA-seq data This study GEO: GSE116580 30 RACE seq data This study GEO: GSE116580 Experimental Models: Cell Lines Mouse: Mouse Embryonic fibroblasts Smug1+/+ Lab made Alsøe et al., 2017 Mouse: Mouse Embryonic fibroblasts Smug1 / Lab made Alsøe et al., 2017 Mouse: Mouse Embryonic fibroblasts Smug1 / _SMUG1 Lab made N/A Mouse: Mouse Embryonic fibroblasts Smug1 / _H241L Lab made N/A Mouse: Mouse Embryonic fibroblasts Smug1 / _E31R/E35R Lab made N/A Human: HAP1 SMUG1-WT (control wild type cell line) Horizon Cat# HZGHC003300c009 Human: HAP1 SMUG1-KO Horizon Cat# HZGHC003300c009 Human: HAP1 SMUG1-KO_SMUG1 Lab made N/A Human: HAP1 SMUG1-KO_SMUG1 H239L (NABm) Lab made N/A Human: HAP1 SMUG1-KO_E29R/E33R/E231R (DBm) Lab made N/A (Continued on next page) Cell Reports 28, 1690–1702.e1–e10, August 13, 2019 e3 .. REAGENT or RESOURCE SOURCE IDENTIFIER Human: HeLa ATCC ATCC CCL-2 Experimental Models: Organisms/Strains C56BL/6 Smug1tm1Hln (Smug1 / ) Lab made Alsøe et al., 2017 Oligonucleotides See Table S1 for oligonucleotide information N/A Recombinant DNA Plasmid: pHH25-Smug1 Cloned and stored in lab N/A Plasmid: pHH25-Smug1 H241L Cloned and stored in lab N/A Plasmid: pHH25-Smug1 E31R/E35R Cloned and stored in lab N/A Plasmid: pHH25-SMUG1 Cloned and stored in lab N/A Plasmid: pHH25-SMUG1 H239L (NABm) Cloned and stored in lab N/A Plasmid: pHH25-SMUG1 E29R/E31R/E231R (DBm) Cloned and stored in lab N/A Plasmid: pCDNA-3xHA-hTERT Addgene Cat# 51631; RRID:Addgene_51637 Plasmid: pBS U3-hTR-500 Addgene Cat# 28170; RRID:Addgene_28170 Plasmid: pETM-11-hSMUG1 wt Cloned and stored in lab N/A Plasmid: pETM-11-hSMUG1 H239L Cloned and stored in lab N/A Software and Algorithms Multi Gauge V3.1 software Fujifilm N/A STAR aligner Dobin et al., 2013 https://github.com/alexdobin/STAR; RRID:SCR_015899 htseq-count Anders et al., 2015 https://github.com/simon-anders/htseq; RRID:SCR_011867 Voom Law et al., 2014 http://bioinf.wehi.edu.au/voom/ Limma Ritchie et al., 2015 https://bioconductor.org/packages/ release/bioc/html/limma.html; RRID:SCR_010943 ImageJ https://imagej.net/Welcome; RRID:SCR_003070 bowtie2 Langmead and Salzberg, 2012 http://bowtie-bio.sourceforge.net/bowtie2/ index.shtml Bedtools Quinlan and Hall, 2010 https://github.com/arq5x/bedtools2; RRID:SCR_006646 TeloTool Göhring et al., 2014 https://github.com/jagoehring/TeloTool Zeiss Zen blue and black Zeiss https://www.zeiss.com/microscopy/en_us/ products/microscope-software/zen.html; RRID:SCR_013672 GraphPad Prism GraphPad https://www.graphpad.com/; RRID:SCR_002798 EZLogic Integration Bio-Rad http://www.bio-rad.com
End Labeling:Article Title: SMUG1 Promotes Telomere Maintenance through Telomerase RNA Processing.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER mouse IL-6 recombinant protein eBioscience Cat# 14-8061-80 mouse stem cell factor (SCF) recombinant protein eBioscience Cat# 14-8341-63 Mouse Methylcellulose Complete Media R&D systems Cat# HSC007 Colcemid Life Technologies Cat# 15210040 Blocking reagent Sigma-Aldrich-Aldrich Cat# 11096176001 Vectashield mounting medium with DAPI Vector labs Cat# H-1200 Pierce BCA Protein Assay Kit Life Technologies Cat# 23227 DNeasy Blood & Tissue Kit QIAGEN Cat# 69504 APE1 New England Biolabs Cat# M0282S Thymidine Sigma-Aldrich-Aldrich Cat# T1895 Imidazole buffer Solution Sigma-Aldrich-Aldrich Cat# 68268 Isopropyl b-D-1-thiogalactopyranoside Sigma-Aldrich-Aldrich Cat# I5502 Ni-NTA agarose resin QIAGEN Cat# 30210 Econo-Column Chromatography Columns Bio-Rad Cat# 7372522 Bio-Scale Mini Macro-Prep High S cartridge Bio-Rad Cat# 7324134 Ultrafree-MC Centrifugal Filter Millipore Cat# UFC30DV0S HIS-Select Nickel Magnetic Agarose beads Sigma-Aldrich-Aldrich Cat# H9914 SMUG1 wt recombinant protein Lab made N/A SMUG1 H239L recombinant protein Lab made N/A Critical Commercial Assays SENSE Total RNA-Seq Library Prep Kit Lexogen Cat# SKU: 009.24 Click-iT Nascent RNA Capture Kit, for gene expression analysis Life Technologies Cat# C10365 DIG Oligonucleotide 30 End Labeling Kit, 2nd generation Sigma-Aldrich-Aldrich Cat# 03353575910 TeloTAGGG Telomere Length Assay Sigma-Aldrich-Aldrich Cat# 12209136001 TruSeq Nano DNA Low Throughput Library Prep Kit Illumina Cat# 20015964 MiSeq Reagent Kit v2 (500-cycles) Illumina Cat# MS-102-2003 Duolink In Situ Red Starter Kit Mouse/Rabbit Sigma-Aldrich-Aldrich Cat# DUO92101 TRAPeze Telomerase Detection Kit Millipore Cat# S7710 MEGAscript T7 Transcription Kit Life Technologies Cat# AM1333 Deposited Data RNA-seq data This study GEO: GSE116580 30 RACE seq data This study GEO: GSE116580 Experimental Models: Cell Lines Mouse: Mouse Embryonic fibroblasts Smug1+/+ Lab made Alsøe et al., 2017 Mouse: Mouse Embryonic fibroblasts Smug1 / Lab made Alsøe et al., 2017 Mouse: Mouse Embryonic fibroblasts Smug1 / _SMUG1 Lab made N/A Mouse: Mouse Embryonic fibroblasts Smug1 / _H241L Lab made N/A Mouse: Mouse Embryonic fibroblasts Smug1 / _E31R/E35R Lab made N/A Human: HAP1 SMUG1-WT (control wild type cell line) Horizon Cat# HZGHC003300c009 Human: HAP1 SMUG1-KO Horizon Cat# HZGHC003300c009 Human: HAP1 SMUG1-KO_SMUG1 Lab made N/A Human: HAP1 SMUG1-KO_SMUG1 H239L (NABm) Lab made N/A Human: HAP1 SMUG1-KO_E29R/E33R/E231R (DBm) Lab made N/A (Continued on next page) Cell Reports 28, 1690–1702.e1–e10, August 13, 2019 e3 .. REAGENT or RESOURCE SOURCE IDENTIFIER Human: HeLa ATCC ATCC CCL-2 Experimental Models: Organisms/Strains C56BL/6 Smug1tm1Hln (Smug1 / ) Lab made Alsøe et al., 2017 Oligonucleotides See Table S1 for oligonucleotide information N/A Recombinant DNA Plasmid: pHH25-Smug1 Cloned and stored in lab N/A Plasmid: pHH25-Smug1 H241L Cloned and stored in lab N/A Plasmid: pHH25-Smug1 E31R/E35R Cloned and stored in lab N/A Plasmid: pHH25-SMUG1 Cloned and stored in lab N/A Plasmid: pHH25-SMUG1 H239L (NABm) Cloned and stored in lab N/A Plasmid: pHH25-SMUG1 E29R/E31R/E231R (DBm) Cloned and stored in lab N/A Plasmid: pCDNA-3xHA-hTERT Addgene Cat# 51631; RRID:Addgene_51637 Plasmid: pBS U3-hTR-500 Addgene Cat# 28170; RRID:Addgene_28170 Plasmid: pETM-11-hSMUG1 wt Cloned and stored in lab N/A Plasmid: pETM-11-hSMUG1 H239L Cloned and stored in lab N/A Software and Algorithms Multi Gauge V3.1 software Fujifilm N/A STAR aligner Dobin et al., 2013 https://github.com/alexdobin/STAR; RRID:SCR_015899 htseq-count Anders et al., 2015 https://github.com/simon-anders/htseq; RRID:SCR_011867 Voom Law et al., 2014 http://bioinf.wehi.edu.au/voom/ Limma Ritchie et al., 2015 https://bioconductor.org/packages/ release/bioc/html/limma.html; RRID:SCR_010943 ImageJ https://imagej.net/Welcome; RRID:SCR_003070 bowtie2 Langmead and Salzberg, 2012 http://bowtie-bio.sourceforge.net/bowtie2/ index.shtml Bedtools Quinlan and Hall, 2010 https://github.com/arq5x/bedtools2; RRID:SCR_006646 TeloTool Göhring et al., 2014 https://github.com/jagoehring/TeloTool Zeiss Zen blue and black Zeiss https://www.zeiss.com/microscopy/en_us/ products/microscope-software/zen.html; RRID:SCR_013672 GraphPad Prism GraphPad https://www.graphpad.com/; RRID:SCR_002798 EZLogic Integration Bio-Rad http://www.bio-rad.com
In Situ:Article Title: SMUG1 Promotes Telomere Maintenance through Telomerase RNA Processing.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER mouse IL-6 recombinant protein eBioscience Cat# 14-8061-80 mouse stem cell factor (SCF) recombinant protein eBioscience Cat# 14-8341-63 Mouse Methylcellulose Complete Media R&D systems Cat# HSC007 Colcemid Life Technologies Cat# 15210040 Blocking reagent Sigma-Aldrich-Aldrich Cat# 11096176001 Vectashield mounting medium with DAPI Vector labs Cat# H-1200 Pierce BCA Protein Assay Kit Life Technologies Cat# 23227 DNeasy Blood & Tissue Kit QIAGEN Cat# 69504 APE1 New England Biolabs Cat# M0282S Thymidine Sigma-Aldrich-Aldrich Cat# T1895 Imidazole buffer Solution Sigma-Aldrich-Aldrich Cat# 68268 Isopropyl b-D-1-thiogalactopyranoside Sigma-Aldrich-Aldrich Cat# I5502 Ni-NTA agarose resin QIAGEN Cat# 30210 Econo-Column Chromatography Columns Bio-Rad Cat# 7372522 Bio-Scale Mini Macro-Prep High S cartridge Bio-Rad Cat# 7324134 Ultrafree-MC Centrifugal Filter Millipore Cat# UFC30DV0S HIS-Select Nickel Magnetic Agarose beads Sigma-Aldrich-Aldrich Cat# H9914 SMUG1 wt recombinant protein Lab made N/A SMUG1 H239L recombinant protein Lab made N/A Critical Commercial Assays SENSE Total RNA-Seq Library Prep Kit Lexogen Cat# SKU: 009.24 Click-iT Nascent RNA Capture Kit, for gene expression analysis Life Technologies Cat# C10365 DIG Oligonucleotide 30 End Labeling Kit, 2nd generation Sigma-Aldrich-Aldrich Cat# 03353575910 TeloTAGGG Telomere Length Assay Sigma-Aldrich-Aldrich Cat# 12209136001 TruSeq Nano DNA Low Throughput Library Prep Kit Illumina Cat# 20015964 MiSeq Reagent Kit v2 (500-cycles) Illumina Cat# MS-102-2003 Duolink In Situ Red Starter Kit Mouse/Rabbit Sigma-Aldrich-Aldrich Cat# DUO92101 TRAPeze Telomerase Detection Kit Millipore Cat# S7710 MEGAscript T7 Transcription Kit Life Technologies Cat# AM1333 Deposited Data RNA-seq data This study GEO: GSE116580 30 RACE seq data This study GEO: GSE116580 Experimental Models: Cell Lines Mouse: Mouse Embryonic fibroblasts Smug1+/+ Lab made Alsøe et al., 2017 Mouse: Mouse Embryonic fibroblasts Smug1 / Lab made Alsøe et al., 2017 Mouse: Mouse Embryonic fibroblasts Smug1 / _SMUG1 Lab made N/A Mouse: Mouse Embryonic fibroblasts Smug1 / _H241L Lab made N/A Mouse: Mouse Embryonic fibroblasts Smug1 / _E31R/E35R Lab made N/A Human: HAP1 SMUG1-WT (control wild type cell line) Horizon Cat# HZGHC003300c009 Human: HAP1 SMUG1-KO Horizon Cat# HZGHC003300c009 Human: HAP1 SMUG1-KO_SMUG1 Lab made N/A Human: HAP1 SMUG1-KO_SMUG1 H239L (NABm) Lab made N/A Human: HAP1 SMUG1-KO_E29R/E33R/E231R (DBm) Lab made N/A (Continued on next page) Cell Reports 28, 1690–1702.e1–e10, August 13, 2019 e3 .. REAGENT or RESOURCE SOURCE IDENTIFIER Human: HeLa ATCC ATCC CCL-2 Experimental Models: Organisms/Strains C56BL/6 Smug1tm1Hln (Smug1 / ) Lab made Alsøe et al., 2017 Oligonucleotides See Table S1 for oligonucleotide information N/A Recombinant DNA Plasmid: pHH25-Smug1 Cloned and stored in lab N/A Plasmid: pHH25-Smug1 H241L Cloned and stored in lab N/A Plasmid: pHH25-Smug1 E31R/E35R Cloned and stored in lab N/A Plasmid: pHH25-SMUG1 Cloned and stored in lab N/A Plasmid: pHH25-SMUG1 H239L (NABm) Cloned and stored in lab N/A Plasmid: pHH25-SMUG1 E29R/E31R/E231R (DBm) Cloned and stored in lab N/A Plasmid: pCDNA-3xHA-hTERT Addgene Cat# 51631; RRID:Addgene_51637 Plasmid: pBS U3-hTR-500 Addgene Cat# 28170; RRID:Addgene_28170 Plasmid: pETM-11-hSMUG1 wt Cloned and stored in lab N/A Plasmid: pETM-11-hSMUG1 H239L Cloned and stored in lab N/A Software and Algorithms Multi Gauge V3.1 software Fujifilm N/A STAR aligner Dobin et al., 2013 https://github.com/alexdobin/STAR; RRID:SCR_015899 htseq-count Anders et al., 2015 https://github.com/simon-anders/htseq; RRID:SCR_011867 Voom Law et al., 2014 http://bioinf.wehi.edu.au/voom/ Limma Ritchie et al., 2015 https://bioconductor.org/packages/ release/bioc/html/limma.html; RRID:SCR_010943 ImageJ https://imagej.net/Welcome; RRID:SCR_003070 bowtie2 Langmead and Salzberg, 2012 http://bowtie-bio.sourceforge.net/bowtie2/ index.shtml Bedtools Quinlan and Hall, 2010 https://github.com/arq5x/bedtools2; RRID:SCR_006646 TeloTool Göhring et al., 2014 https://github.com/jagoehring/TeloTool Zeiss Zen blue and black Zeiss https://www.zeiss.com/microscopy/en_us/ products/microscope-software/zen.html; RRID:SCR_013672 GraphPad Prism GraphPad https://www.graphpad.com/; RRID:SCR_002798 EZLogic Integration Bio-Rad http://www.bio-rad.com
RNA Sequencing:Article Title: SMUG1 Promotes Telomere Maintenance through Telomerase RNA Processing.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER mouse IL-6 recombinant protein eBioscience Cat# 14-8061-80 mouse stem cell factor (SCF) recombinant protein eBioscience Cat# 14-8341-63 Mouse Methylcellulose Complete Media R&D systems Cat# HSC007 Colcemid Life Technologies Cat# 15210040 Blocking reagent Sigma-Aldrich-Aldrich Cat# 11096176001 Vectashield mounting medium with DAPI Vector labs Cat# H-1200 Pierce BCA Protein Assay Kit Life Technologies Cat# 23227 DNeasy Blood & Tissue Kit QIAGEN Cat# 69504 APE1 New England Biolabs Cat# M0282S Thymidine Sigma-Aldrich-Aldrich Cat# T1895 Imidazole buffer Solution Sigma-Aldrich-Aldrich Cat# 68268 Isopropyl b-D-1-thiogalactopyranoside Sigma-Aldrich-Aldrich Cat# I5502 Ni-NTA agarose resin QIAGEN Cat# 30210 Econo-Column Chromatography Columns Bio-Rad Cat# 7372522 Bio-Scale Mini Macro-Prep High S cartridge Bio-Rad Cat# 7324134 Ultrafree-MC Centrifugal Filter Millipore Cat# UFC30DV0S HIS-Select Nickel Magnetic Agarose beads Sigma-Aldrich-Aldrich Cat# H9914 SMUG1 wt recombinant protein Lab made N/A SMUG1 H239L recombinant protein Lab made N/A Critical Commercial Assays SENSE Total RNA-Seq Library Prep Kit Lexogen Cat# SKU: 009.24 Click-iT Nascent RNA Capture Kit, for gene expression analysis Life Technologies Cat# C10365 DIG Oligonucleotide 30 End Labeling Kit, 2nd generation Sigma-Aldrich-Aldrich Cat# 03353575910 TeloTAGGG Telomere Length Assay Sigma-Aldrich-Aldrich Cat# 12209136001 TruSeq Nano DNA Low Throughput Library Prep Kit Illumina Cat# 20015964 MiSeq Reagent Kit v2 (500-cycles) Illumina Cat# MS-102-2003 Duolink In Situ Red Starter Kit Mouse/Rabbit Sigma-Aldrich-Aldrich Cat# DUO92101 TRAPeze Telomerase Detection Kit Millipore Cat# S7710 MEGAscript T7 Transcription Kit Life Technologies Cat# AM1333 Deposited Data RNA-seq data This study GEO: GSE116580 30 RACE seq data This study GEO: GSE116580 Experimental Models: Cell Lines Mouse: Mouse Embryonic fibroblasts Smug1+/+ Lab made Alsøe et al., 2017 Mouse: Mouse Embryonic fibroblasts Smug1 / Lab made Alsøe et al., 2017 Mouse: Mouse Embryonic fibroblasts Smug1 / _SMUG1 Lab made N/A Mouse: Mouse Embryonic fibroblasts Smug1 / _H241L Lab made N/A Mouse: Mouse Embryonic fibroblasts Smug1 / _E31R/E35R Lab made N/A Human: HAP1 SMUG1-WT (control wild type cell line) Horizon Cat# HZGHC003300c009 Human: HAP1 SMUG1-KO Horizon Cat# HZGHC003300c009 Human: HAP1 SMUG1-KO_SMUG1 Lab made N/A Human: HAP1 SMUG1-KO_SMUG1 H239L (NABm) Lab made N/A Human: HAP1 SMUG1-KO_E29R/E33R/E231R (DBm) Lab made N/A (Continued on next page) Cell Reports 28, 1690–1702.e1–e10, August 13, 2019 e3 .. REAGENT or RESOURCE SOURCE IDENTIFIER Human: HeLa ATCC ATCC CCL-2 Experimental Models: Organisms/Strains C56BL/6 Smug1tm1Hln (Smug1 / ) Lab made Alsøe et al., 2017 Oligonucleotides See Table S1 for oligonucleotide information N/A Recombinant DNA Plasmid: pHH25-Smug1 Cloned and stored in lab N/A Plasmid: pHH25-Smug1 H241L Cloned and stored in lab N/A Plasmid: pHH25-Smug1 E31R/E35R Cloned and stored in lab N/A Plasmid: pHH25-SMUG1 Cloned and stored in lab N/A Plasmid: pHH25-SMUG1 H239L (NABm) Cloned and stored in lab N/A Plasmid: pHH25-SMUG1 E29R/E31R/E231R (DBm) Cloned and stored in lab N/A Plasmid: pCDNA-3xHA-hTERT Addgene Cat# 51631; RRID:Addgene_51637 Plasmid: pBS U3-hTR-500 Addgene Cat# 28170; RRID:Addgene_28170 Plasmid: pETM-11-hSMUG1 wt Cloned and stored in lab N/A Plasmid: pETM-11-hSMUG1 H239L Cloned and stored in lab N/A Software and Algorithms Multi Gauge V3.1 software Fujifilm N/A STAR aligner Dobin et al., 2013 https://github.com/alexdobin/STAR; RRID:SCR_015899 htseq-count Anders et al., 2015 https://github.com/simon-anders/htseq; RRID:SCR_011867 Voom Law et al., 2014 http://bioinf.wehi.edu.au/voom/ Limma Ritchie et al., 2015 https://bioconductor.org/packages/ release/bioc/html/limma.html; RRID:SCR_010943 ImageJ https://imagej.net/Welcome; RRID:SCR_003070 bowtie2 Langmead and Salzberg, 2012 http://bowtie-bio.sourceforge.net/bowtie2/ index.shtml Bedtools Quinlan and Hall, 2010 https://github.com/arq5x/bedtools2; RRID:SCR_006646 TeloTool Göhring et al., 2014 https://github.com/jagoehring/TeloTool Zeiss Zen blue and black Zeiss https://www.zeiss.com/microscopy/en_us/ products/microscope-software/zen.html; RRID:SCR_013672 GraphPad Prism GraphPad https://www.graphpad.com/; RRID:SCR_002798 EZLogic Integration Bio-Rad http://www.bio-rad.com
Control:Article Title: SMUG1 Promotes Telomere Maintenance through Telomerase RNA Processing.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER mouse IL-6 recombinant protein eBioscience Cat# 14-8061-80 mouse stem cell factor (SCF) recombinant protein eBioscience Cat# 14-8341-63 Mouse Methylcellulose Complete Media R&D systems Cat# HSC007 Colcemid Life Technologies Cat# 15210040 Blocking reagent Sigma-Aldrich-Aldrich Cat# 11096176001 Vectashield mounting medium with DAPI Vector labs Cat# H-1200 Pierce BCA Protein Assay Kit Life Technologies Cat# 23227 DNeasy Blood & Tissue Kit QIAGEN Cat# 69504 APE1 New England Biolabs Cat# M0282S Thymidine Sigma-Aldrich-Aldrich Cat# T1895 Imidazole buffer Solution Sigma-Aldrich-Aldrich Cat# 68268 Isopropyl b-D-1-thiogalactopyranoside Sigma-Aldrich-Aldrich Cat# I5502 Ni-NTA agarose resin QIAGEN Cat# 30210 Econo-Column Chromatography Columns Bio-Rad Cat# 7372522 Bio-Scale Mini Macro-Prep High S cartridge Bio-Rad Cat# 7324134 Ultrafree-MC Centrifugal Filter Millipore Cat# UFC30DV0S HIS-Select Nickel Magnetic Agarose beads Sigma-Aldrich-Aldrich Cat# H9914 SMUG1 wt recombinant protein Lab made N/A SMUG1 H239L recombinant protein Lab made N/A Critical Commercial Assays SENSE Total RNA-Seq Library Prep Kit Lexogen Cat# SKU: 009.24 Click-iT Nascent RNA Capture Kit, for gene expression analysis Life Technologies Cat# C10365 DIG Oligonucleotide 30 End Labeling Kit, 2nd generation Sigma-Aldrich-Aldrich Cat# 03353575910 TeloTAGGG Telomere Length Assay Sigma-Aldrich-Aldrich Cat# 12209136001 TruSeq Nano DNA Low Throughput Library Prep Kit Illumina Cat# 20015964 MiSeq Reagent Kit v2 (500-cycles) Illumina Cat# MS-102-2003 Duolink In Situ Red Starter Kit Mouse/Rabbit Sigma-Aldrich-Aldrich Cat# DUO92101 TRAPeze Telomerase Detection Kit Millipore Cat# S7710 MEGAscript T7 Transcription Kit Life Technologies Cat# AM1333 Deposited Data RNA-seq data This study GEO: GSE116580 30 RACE seq data This study GEO: GSE116580 Experimental Models: Cell Lines Mouse: Mouse Embryonic fibroblasts Smug1+/+ Lab made Alsøe et al., 2017 Mouse: Mouse Embryonic fibroblasts Smug1 / Lab made Alsøe et al., 2017 Mouse: Mouse Embryonic fibroblasts Smug1 / _SMUG1 Lab made N/A Mouse: Mouse Embryonic fibroblasts Smug1 / _H241L Lab made N/A Mouse: Mouse Embryonic fibroblasts Smug1 / _E31R/E35R Lab made N/A Human: HAP1 SMUG1-WT (control wild type cell line) Horizon Cat# HZGHC003300c009 Human: HAP1 SMUG1-KO Horizon Cat# HZGHC003300c009 Human: HAP1 SMUG1-KO_SMUG1 Lab made N/A Human: HAP1 SMUG1-KO_SMUG1 H239L (NABm) Lab made N/A Human: HAP1 SMUG1-KO_E29R/E33R/E231R (DBm) Lab made N/A (Continued on next page) Cell Reports 28, 1690–1702.e1–e10, August 13, 2019 e3 .. REAGENT or RESOURCE SOURCE IDENTIFIER Human: HeLa ATCC ATCC CCL-2 Experimental Models: Organisms/Strains C56BL/6 Smug1tm1Hln (Smug1 / ) Lab made Alsøe et al., 2017 Oligonucleotides See Table S1 for oligonucleotide information N/A Recombinant DNA Plasmid: pHH25-Smug1 Cloned and stored in lab N/A Plasmid: pHH25-Smug1 H241L Cloned and stored in lab N/A Plasmid: pHH25-Smug1 E31R/E35R Cloned and stored in lab N/A Plasmid: pHH25-SMUG1 Cloned and stored in lab N/A Plasmid: pHH25-SMUG1 H239L (NABm) Cloned and stored in lab N/A Plasmid: pHH25-SMUG1 E29R/E31R/E231R (DBm) Cloned and stored in lab N/A Plasmid: pCDNA-3xHA-hTERT Addgene Cat# 51631; RRID:Addgene_51637 Plasmid: pBS U3-hTR-500 Addgene Cat# 28170; RRID:Addgene_28170 Plasmid: pETM-11-hSMUG1 wt Cloned and stored in lab N/A Plasmid: pETM-11-hSMUG1 H239L Cloned and stored in lab N/A Software and Algorithms Multi Gauge V3.1 software Fujifilm N/A STAR aligner Dobin et al., 2013 https://github.com/alexdobin/STAR; RRID:SCR_015899 htseq-count Anders et al., 2015 https://github.com/simon-anders/htseq; RRID:SCR_011867 Voom Law et al., 2014 http://bioinf.wehi.edu.au/voom/ Limma Ritchie et al., 2015 https://bioconductor.org/packages/ release/bioc/html/limma.html; RRID:SCR_010943 ImageJ https://imagej.net/Welcome; RRID:SCR_003070 bowtie2 Langmead and Salzberg, 2012 http://bowtie-bio.sourceforge.net/bowtie2/ index.shtml Bedtools Quinlan and Hall, 2010 https://github.com/arq5x/bedtools2; RRID:SCR_006646 TeloTool Göhring et al., 2014 https://github.com/jagoehring/TeloTool Zeiss Zen blue and black Zeiss https://www.zeiss.com/microscopy/en_us/ products/microscope-software/zen.html; RRID:SCR_013672 GraphPad Prism GraphPad https://www.graphpad.com/; RRID:SCR_002798 EZLogic Integration Bio-Rad http://www.bio-rad.com
other:Article Title: SMUG1 Promotes Telomere Maintenance through Telomerase RNA Processing.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Human: HeLa ATCC ATCC CCL-2 Experimental Models: Organisms/Strains C56BL/6 Smug1tm1Hln (Smug1 / ) Lab made Alsøe et al., 2017 Oligonucleotides See Table S1 for oligonucleotide information N/A Recombinant DNA Plasmid: pHH25-Smug1 Cloned and stored in lab N/A Plasmid: pHH25-Smug1 H241L Cloned and stored in lab N/A Plasmid: pHH25-Smug1 E31R/E35R Cloned and stored in lab N/A Plasmid: pHH25-SMUG1 Cloned and stored in lab N/A Plasmid: pHH25-SMUG1 H239L (NABm) Cloned and stored in lab N/A Plasmid: pHH25-SMUG1 E29R/E31R/E231R (DBm) Cloned and stored in lab N/A Plasmid: pCDNA-3xHA-hTERT Addgene Cat# 51631; RRID:Addgene_51637 Plasmid: pBS U3-hTR-500 Addgene Cat# 28170; RRID:Addgene_28170 Plasmid: pETM-11-hSMUG1 wt Cloned and stored in lab N/A Plasmid: pETM-11-hSMUG1 H239L Cloned and stored in lab N/A Software and Algorithms Multi Gauge V3.1 software Fujifilm N/A STAR aligner Dobin et al., 2013 https://github.com/alexdobin/STAR; RRID:SCR_015899 htseq-count Anders et al., 2015 https://github.com/simon-anders/htseq; RRID:SCR_011867 Voom Law et al., 2014 http://bioinf.wehi.edu.au/voom/ Limma Ritchie et al., 2015 https://bioconductor.org/packages/ release/bioc/html/limma.html; RRID:SCR_010943 ImageJ https://imagej.net/Welcome; RRID:SCR_003070 bowtie2 Langmead and Salzberg, 2012 http://bowtie-bio.sourceforge.net/bowtie2/ index.shtml Bedtools Quinlan and Hall, 2010 https://github.com/arq5x/bedtools2; RRID:SCR_006646 TeloTool Göhring et al., 2014 https://github.com/jagoehring/TeloTool Zeiss Zen blue and black Zeiss https://www.zeiss.com/microscopy/en_us/ products/microscope-software/zen.html; RRID:SCR_013672 GraphPad Prism GraphPad https://www.graphpad.com/; RRID:SCR_002798 EZLogic Integration Bio-Rad http://www.bio-rad.com
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