Plasmid Preparation:Article Title: TRF1 and TRF2 form distinct shelterin subcomplexes at telomeres.
Article Snippet: All generated HeLa cell lines were based on HeLa-EM2-11ht 50 and were grown in DMEM media (Gibco) supplemented with 10% FBS, 100 U ml − 1 penicillin and 100 μg mL − 1 streptomycin (Gibco) at 37 ◦ C and 5% CO 2 . .. REAGENT or RESOURCE SOURCE IDENTIFIER pX330-TIN2 plasmid This study N/A pX330-TPP1 plasmid This study N/A pX330-POT1 plasmid This study N/A pX330-RAP1 plasmid This study N/A eSpCas9-ATP1A1-G2-Dual-sgRNA plasmid Agudelo et al. 82 Addgene plasmid #86612 eSpCas9-ATP1A1-G2-Dual-RAP1-KO plasmid This study N/A 6xHis-3xFLAG-HaloTag plasmid Thomas R Cech N/A pHTN HaloTag CMV-neo plasmid Promega Cat#G7721 Software and algorithms Fiji 2.16.0 Schindelin et al. 83 https://imagej.net/software/fiji/ MATLAB 2024b Mathworks Inc. https://www.mathworks.com/ Python 3.11.5 Python Software Foundation https://www.python.org Image Quant TL 8.2 Cytiva https://www.cytivalifesciences.com/en/us/ shop/protein-analysis/molecular-imagingfor-proteins/imaging-software/ imagequant-tl-analysis-software-p-28619 FCS Express 7 Research De Novo Software https://denovosoftware.com/ SpectroFlo v2.2 Cytec https://cytekbio.com/pages/spectro-flo R Studio 2022.07.2 Posit Software, PBC https://posit.co/products/open-source/ rstudio/ GraphPad Prism 10.5.0 GraphPad Software https://www.graphpad.com/ SPT LocAndTrack algorithm Hansen et al. 59 https://github.com/elifesciences- publications/SPT_LocAndTrack Spot-On algorithm Hansen et al. 59 https://gitlab.com/tjian-darzacq-lab/spot- on-matlab ExTrack algorithm Simon et al. 60 https://github.com/vanTeeffelenLab/ ExTrack Cellpose 2.0 segmentation algorithm Stringer et al. 84 https://github.com/MouseLand/cellpose ComDet plugin for Fiji v.0.5.5 Katrukha et al. 85 https://imagej.net/plugins/spots- colocalization-comdet Photobleaching and residence time analysis algorithms This study https://github.com/SchmidtLabMSU/ ShelterinAnalysis 18 Cell Reports 44, 116178, September 23, 2025 Generation of HaloTagged cell lines and plasmid construction for genome editing To insert HaloTag at the 5 ′ end of all shelterin genes, the CRISPR/Cas9 genome-editing strategy was used with following procedure. .. All sgRNAs were cloned into BpiI-digested pX330-U6-Chimeric-BB-CBh-hSpCas9 backbone (Addgene #42230, gift from Feng Zhang).
Software:Article Title: TRF1 and TRF2 form distinct shelterin subcomplexes at telomeres.
Article Snippet: All generated HeLa cell lines were based on HeLa-EM2-11ht 50 and were grown in DMEM media (Gibco) supplemented with 10% FBS, 100 U ml − 1 penicillin and 100 μg mL − 1 streptomycin (Gibco) at 37 ◦ C and 5% CO 2 . .. REAGENT or RESOURCE SOURCE IDENTIFIER pX330-TIN2 plasmid This study N/A pX330-TPP1 plasmid This study N/A pX330-POT1 plasmid This study N/A pX330-RAP1 plasmid This study N/A eSpCas9-ATP1A1-G2-Dual-sgRNA plasmid Agudelo et al. 82 Addgene plasmid #86612 eSpCas9-ATP1A1-G2-Dual-RAP1-KO plasmid This study N/A 6xHis-3xFLAG-HaloTag plasmid Thomas R Cech N/A pHTN HaloTag CMV-neo plasmid Promega Cat#G7721 Software and algorithms Fiji 2.16.0 Schindelin et al. 83 https://imagej.net/software/fiji/ MATLAB 2024b Mathworks Inc. https://www.mathworks.com/ Python 3.11.5 Python Software Foundation https://www.python.org Image Quant TL 8.2 Cytiva https://www.cytivalifesciences.com/en/us/ shop/protein-analysis/molecular-imagingfor-proteins/imaging-software/ imagequant-tl-analysis-software-p-28619 FCS Express 7 Research De Novo Software https://denovosoftware.com/ SpectroFlo v2.2 Cytec https://cytekbio.com/pages/spectro-flo R Studio 2022.07.2 Posit Software, PBC https://posit.co/products/open-source/ rstudio/ GraphPad Prism 10.5.0 GraphPad Software https://www.graphpad.com/ SPT LocAndTrack algorithm Hansen et al. 59 https://github.com/elifesciences- publications/SPT_LocAndTrack Spot-On algorithm Hansen et al. 59 https://gitlab.com/tjian-darzacq-lab/spot- on-matlab ExTrack algorithm Simon et al. 60 https://github.com/vanTeeffelenLab/ ExTrack Cellpose 2.0 segmentation algorithm Stringer et al. 84 https://github.com/MouseLand/cellpose ComDet plugin for Fiji v.0.5.5 Katrukha et al. 85 https://imagej.net/plugins/spots- colocalization-comdet Photobleaching and residence time analysis algorithms This study https://github.com/SchmidtLabMSU/ ShelterinAnalysis 18 Cell Reports 44, 116178, September 23, 2025 Generation of HaloTagged cell lines and plasmid construction for genome editing To insert HaloTag at the 5 ′ end of all shelterin genes, the CRISPR/Cas9 genome-editing strategy was used with following procedure. .. All sgRNAs were cloned into BpiI-digested pX330-U6-Chimeric-BB-CBh-hSpCas9 backbone (Addgene #42230, gift from Feng Zhang).
Single-particle Tracking:Article Title: TRF1 and TRF2 form distinct shelterin subcomplexes at telomeres.
Article Snippet: All generated HeLa cell lines were based on HeLa-EM2-11ht 50 and were grown in DMEM media (Gibco) supplemented with 10% FBS, 100 U ml − 1 penicillin and 100 μg mL − 1 streptomycin (Gibco) at 37 ◦ C and 5% CO 2 . .. REAGENT or RESOURCE SOURCE IDENTIFIER pX330-TIN2 plasmid This study N/A pX330-TPP1 plasmid This study N/A pX330-POT1 plasmid This study N/A pX330-RAP1 plasmid This study N/A eSpCas9-ATP1A1-G2-Dual-sgRNA plasmid Agudelo et al. 82 Addgene plasmid #86612 eSpCas9-ATP1A1-G2-Dual-RAP1-KO plasmid This study N/A 6xHis-3xFLAG-HaloTag plasmid Thomas R Cech N/A pHTN HaloTag CMV-neo plasmid Promega Cat#G7721 Software and algorithms Fiji 2.16.0 Schindelin et al. 83 https://imagej.net/software/fiji/ MATLAB 2024b Mathworks Inc. https://www.mathworks.com/ Python 3.11.5 Python Software Foundation https://www.python.org Image Quant TL 8.2 Cytiva https://www.cytivalifesciences.com/en/us/ shop/protein-analysis/molecular-imagingfor-proteins/imaging-software/ imagequant-tl-analysis-software-p-28619 FCS Express 7 Research De Novo Software https://denovosoftware.com/ SpectroFlo v2.2 Cytec https://cytekbio.com/pages/spectro-flo R Studio 2022.07.2 Posit Software, PBC https://posit.co/products/open-source/ rstudio/ GraphPad Prism 10.5.0 GraphPad Software https://www.graphpad.com/ SPT LocAndTrack algorithm Hansen et al. 59 https://github.com/elifesciences- publications/SPT_LocAndTrack Spot-On algorithm Hansen et al. 59 https://gitlab.com/tjian-darzacq-lab/spot- on-matlab ExTrack algorithm Simon et al. 60 https://github.com/vanTeeffelenLab/ ExTrack Cellpose 2.0 segmentation algorithm Stringer et al. 84 https://github.com/MouseLand/cellpose ComDet plugin for Fiji v.0.5.5 Katrukha et al. 85 https://imagej.net/plugins/spots- colocalization-comdet Photobleaching and residence time analysis algorithms This study https://github.com/SchmidtLabMSU/ ShelterinAnalysis 18 Cell Reports 44, 116178, September 23, 2025 Generation of HaloTagged cell lines and plasmid construction for genome editing To insert HaloTag at the 5 ′ end of all shelterin genes, the CRISPR/Cas9 genome-editing strategy was used with following procedure. .. All sgRNAs were cloned into BpiI-digested pX330-U6-Chimeric-BB-CBh-hSpCas9 backbone (Addgene #42230, gift from Feng Zhang).
CRISPR:Article Title: TRF1 and TRF2 form distinct shelterin subcomplexes at telomeres.
Article Snippet: All generated HeLa cell lines were based on HeLa-EM2-11ht 50 and were grown in DMEM media (Gibco) supplemented with 10% FBS, 100 U ml − 1 penicillin and 100 μg mL − 1 streptomycin (Gibco) at 37 ◦ C and 5% CO 2 . .. REAGENT or RESOURCE SOURCE IDENTIFIER pX330-TIN2 plasmid This study N/A pX330-TPP1 plasmid This study N/A pX330-POT1 plasmid This study N/A pX330-RAP1 plasmid This study N/A eSpCas9-ATP1A1-G2-Dual-sgRNA plasmid Agudelo et al. 82 Addgene plasmid #86612 eSpCas9-ATP1A1-G2-Dual-RAP1-KO plasmid This study N/A 6xHis-3xFLAG-HaloTag plasmid Thomas R Cech N/A pHTN HaloTag CMV-neo plasmid Promega Cat#G7721 Software and algorithms Fiji 2.16.0 Schindelin et al. 83 https://imagej.net/software/fiji/ MATLAB 2024b Mathworks Inc. https://www.mathworks.com/ Python 3.11.5 Python Software Foundation https://www.python.org Image Quant TL 8.2 Cytiva https://www.cytivalifesciences.com/en/us/ shop/protein-analysis/molecular-imagingfor-proteins/imaging-software/ imagequant-tl-analysis-software-p-28619 FCS Express 7 Research De Novo Software https://denovosoftware.com/ SpectroFlo v2.2 Cytec https://cytekbio.com/pages/spectro-flo R Studio 2022.07.2 Posit Software, PBC https://posit.co/products/open-source/ rstudio/ GraphPad Prism 10.5.0 GraphPad Software https://www.graphpad.com/ SPT LocAndTrack algorithm Hansen et al. 59 https://github.com/elifesciences- publications/SPT_LocAndTrack Spot-On algorithm Hansen et al. 59 https://gitlab.com/tjian-darzacq-lab/spot- on-matlab ExTrack algorithm Simon et al. 60 https://github.com/vanTeeffelenLab/ ExTrack Cellpose 2.0 segmentation algorithm Stringer et al. 84 https://github.com/MouseLand/cellpose ComDet plugin for Fiji v.0.5.5 Katrukha et al. 85 https://imagej.net/plugins/spots- colocalization-comdet Photobleaching and residence time analysis algorithms This study https://github.com/SchmidtLabMSU/ ShelterinAnalysis 18 Cell Reports 44, 116178, September 23, 2025 Generation of HaloTagged cell lines and plasmid construction for genome editing To insert HaloTag at the 5 ′ end of all shelterin genes, the CRISPR/Cas9 genome-editing strategy was used with following procedure. .. All sgRNAs were cloned into BpiI-digested pX330-U6-Chimeric-BB-CBh-hSpCas9 backbone (Addgene #42230, gift from Feng Zhang).
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