Milk:Article Title: ER stress induced mitochondrial dysfunction drives Treg instability in coronary artery disease.
Article Snippet: Reagents and tools table Reagent/resource Reference or source Identifier or catalog number Experimental models Healthy human PBMC’s Clinical core, La Jolla Institute for Immunology (LJI), La Jolla, California, USA PBMCs from CAD patients University of Virginia Health System, Charlottesville, Virginia, USA Coronary Assessment in Virginia (CAVA) cohort Recombinant DNA Not applicable Antibodies Alexa Fluor® 700 antihuman CD8a, Clone RPAT8 Biolegend Cat.#301028, 1:100 Pacific BlueTM anti-human CD4, Clone RPA-T4 Biolegend Cat. #300524, 1:100 10 EMBO Molecular Medicine © The Author(s) D ow nloaded from https://w w w .em bopress.org on N ovem ber 8, 2025 from IP 182.10.130.72. .. Reagent/resource Reference or source Identifier or catalog number Bovine serum albumin Cell Signaling Cat. #9998S Blotting Grade Blocker Non-Fat Dry Milk Biorad Cat. #1706404XTU TGXTM FastCastTM Acrylamide Kit, 12% Biorad Cat. #1610175 Ammonium persulfate Sigma Cat. #A3678 TEMED Biorad Cat. #1610800 RIPA Lysis buffer Cell Signaling Cat. #9806 Protease Inhibitor Roche Cat.#11697498001 4X Laemmli sample buffer Biorad Cat.#1610747 2-Mercaptoethanol ThermoFisher Cat.#21985023 SuperSignalTM West Femto Maximum Sensitivity Substrate 100ml ThermoFisher Cat.#34095 DynabeadsTM Human T-Activator CD3/CD28 for T Cell Expansion and Activation ThermoFisher Cat.#11161D TexMACS Miltenyi Biotec Cat.#130-097-196 RPMI 1640 Medium, no phenol red ThermoFisher Cat.#11835030 Puromycin Sigma Cat.#P7255 2-Deoxy-D-Glucose Sigma Cat.#D6134 Oligomycin A Sigma Cat.#75351 eBioscienceTM Fixation/ Permeabilization Concentrate ThermoFisher Cat.#00-5123-43 eBioscienceTM Fixation/ Permeabilization Diluent ThermoFisher Cat.#00-5223-56 eBioscienceTM Permeabilization Buffer (10X) ThermoFisher Cat.#00-8333-56 Glass-bottom dishes Cellvis Cat.#D35-20-1.5-N Poly D lysine ThermoFisher Cat.#A3890401 EasySepTM Human CD4+ T Cell Isolation Kit STEMCELL Technologies Cat.#17952 EasySepTM Human Naïve CD4+ T Cell Isolation Kit STEMCELL Technologies Cat.#19555 EasySepTM Human CD4+CD127lowCD25+ Regulatory T Cell Isolation Kit STEMCELL Technologies Cat.#18063 ImmunoCultTM-XF T Cell Expansion Medium STEMCELL Technologies Cat.#10981 ImmunoCultTM Human Treg Differentiation Supplement STEMCELL Technologies Cat.#10977 1×Cell Stimulation Cocktail, eBioscience ThermoFisher Cat.# 00-4970-93 1× Protein Transport Inhibitor Cocktail, eBioscience ThermoFisher Cat.# 00-4980-93 Software FlowJo version 10.10.0. .. Becton Dickinson (BD) Reagent/resource Reference or source Identifier or catalog number FIJI is just ImageJ ImageJ, NIH GraphPad Prism version 10 GraphPad Gene set enrichment analysis UC San Diego/Broad Institute https:// www.gseamsigdb.org/gsea/ index.jsp Other Cytek Aurora 5-Laser Spectral Cytometer Cytek Biosciences Nikon AX R with NSPARC confocal system Nikon
Lysis:Article Title: ER stress induced mitochondrial dysfunction drives Treg instability in coronary artery disease.
Article Snippet: Reagents and tools table Reagent/resource Reference or source Identifier or catalog number Experimental models Healthy human PBMC’s Clinical core, La Jolla Institute for Immunology (LJI), La Jolla, California, USA PBMCs from CAD patients University of Virginia Health System, Charlottesville, Virginia, USA Coronary Assessment in Virginia (CAVA) cohort Recombinant DNA Not applicable Antibodies Alexa Fluor® 700 antihuman CD8a, Clone RPAT8 Biolegend Cat.#301028, 1:100 Pacific BlueTM anti-human CD4, Clone RPA-T4 Biolegend Cat. #300524, 1:100 10 EMBO Molecular Medicine © The Author(s) D ow nloaded from https://w w w .em bopress.org on N ovem ber 8, 2025 from IP 182.10.130.72. .. Reagent/resource Reference or source Identifier or catalog number Bovine serum albumin Cell Signaling Cat. #9998S Blotting Grade Blocker Non-Fat Dry Milk Biorad Cat. #1706404XTU TGXTM FastCastTM Acrylamide Kit, 12% Biorad Cat. #1610175 Ammonium persulfate Sigma Cat. #A3678 TEMED Biorad Cat. #1610800 RIPA Lysis buffer Cell Signaling Cat. #9806 Protease Inhibitor Roche Cat.#11697498001 4X Laemmli sample buffer Biorad Cat.#1610747 2-Mercaptoethanol ThermoFisher Cat.#21985023 SuperSignalTM West Femto Maximum Sensitivity Substrate 100ml ThermoFisher Cat.#34095 DynabeadsTM Human T-Activator CD3/CD28 for T Cell Expansion and Activation ThermoFisher Cat.#11161D TexMACS Miltenyi Biotec Cat.#130-097-196 RPMI 1640 Medium, no phenol red ThermoFisher Cat.#11835030 Puromycin Sigma Cat.#P7255 2-Deoxy-D-Glucose Sigma Cat.#D6134 Oligomycin A Sigma Cat.#75351 eBioscienceTM Fixation/ Permeabilization Concentrate ThermoFisher Cat.#00-5123-43 eBioscienceTM Fixation/ Permeabilization Diluent ThermoFisher Cat.#00-5223-56 eBioscienceTM Permeabilization Buffer (10X) ThermoFisher Cat.#00-8333-56 Glass-bottom dishes Cellvis Cat.#D35-20-1.5-N Poly D lysine ThermoFisher Cat.#A3890401 EasySepTM Human CD4+ T Cell Isolation Kit STEMCELL Technologies Cat.#17952 EasySepTM Human Naïve CD4+ T Cell Isolation Kit STEMCELL Technologies Cat.#19555 EasySepTM Human CD4+CD127lowCD25+ Regulatory T Cell Isolation Kit STEMCELL Technologies Cat.#18063 ImmunoCultTM-XF T Cell Expansion Medium STEMCELL Technologies Cat.#10981 ImmunoCultTM Human Treg Differentiation Supplement STEMCELL Technologies Cat.#10977 1×Cell Stimulation Cocktail, eBioscience ThermoFisher Cat.# 00-4970-93 1× Protein Transport Inhibitor Cocktail, eBioscience ThermoFisher Cat.# 00-4980-93 Software FlowJo version 10.10.0. .. Becton Dickinson (BD) Reagent/resource Reference or source Identifier or catalog number FIJI is just ImageJ ImageJ, NIH GraphPad Prism version 10 GraphPad Gene set enrichment analysis UC San Diego/Broad Institute https:// www.gseamsigdb.org/gsea/ index.jsp Other Cytek Aurora 5-Laser Spectral Cytometer Cytek Biosciences Nikon AX R with NSPARC confocal system Nikon
Protease Inhibitor:Article Title: ER stress induced mitochondrial dysfunction drives Treg instability in coronary artery disease.
Article Snippet: Reagents and tools table Reagent/resource Reference or source Identifier or catalog number Experimental models Healthy human PBMC’s Clinical core, La Jolla Institute for Immunology (LJI), La Jolla, California, USA PBMCs from CAD patients University of Virginia Health System, Charlottesville, Virginia, USA Coronary Assessment in Virginia (CAVA) cohort Recombinant DNA Not applicable Antibodies Alexa Fluor® 700 antihuman CD8a, Clone RPAT8 Biolegend Cat.#301028, 1:100 Pacific BlueTM anti-human CD4, Clone RPA-T4 Biolegend Cat. #300524, 1:100 10 EMBO Molecular Medicine © The Author(s) D ow nloaded from https://w w w .em bopress.org on N ovem ber 8, 2025 from IP 182.10.130.72. .. Reagent/resource Reference or source Identifier or catalog number Bovine serum albumin Cell Signaling Cat. #9998S Blotting Grade Blocker Non-Fat Dry Milk Biorad Cat. #1706404XTU TGXTM FastCastTM Acrylamide Kit, 12% Biorad Cat. #1610175 Ammonium persulfate Sigma Cat. #A3678 TEMED Biorad Cat. #1610800 RIPA Lysis buffer Cell Signaling Cat. #9806 Protease Inhibitor Roche Cat.#11697498001 4X Laemmli sample buffer Biorad Cat.#1610747 2-Mercaptoethanol ThermoFisher Cat.#21985023 SuperSignalTM West Femto Maximum Sensitivity Substrate 100ml ThermoFisher Cat.#34095 DynabeadsTM Human T-Activator CD3/CD28 for T Cell Expansion and Activation ThermoFisher Cat.#11161D TexMACS Miltenyi Biotec Cat.#130-097-196 RPMI 1640 Medium, no phenol red ThermoFisher Cat.#11835030 Puromycin Sigma Cat.#P7255 2-Deoxy-D-Glucose Sigma Cat.#D6134 Oligomycin A Sigma Cat.#75351 eBioscienceTM Fixation/ Permeabilization Concentrate ThermoFisher Cat.#00-5123-43 eBioscienceTM Fixation/ Permeabilization Diluent ThermoFisher Cat.#00-5223-56 eBioscienceTM Permeabilization Buffer (10X) ThermoFisher Cat.#00-8333-56 Glass-bottom dishes Cellvis Cat.#D35-20-1.5-N Poly D lysine ThermoFisher Cat.#A3890401 EasySepTM Human CD4+ T Cell Isolation Kit STEMCELL Technologies Cat.#17952 EasySepTM Human Naïve CD4+ T Cell Isolation Kit STEMCELL Technologies Cat.#19555 EasySepTM Human CD4+CD127lowCD25+ Regulatory T Cell Isolation Kit STEMCELL Technologies Cat.#18063 ImmunoCultTM-XF T Cell Expansion Medium STEMCELL Technologies Cat.#10981 ImmunoCultTM Human Treg Differentiation Supplement STEMCELL Technologies Cat.#10977 1×Cell Stimulation Cocktail, eBioscience ThermoFisher Cat.# 00-4970-93 1× Protein Transport Inhibitor Cocktail, eBioscience ThermoFisher Cat.# 00-4980-93 Software FlowJo version 10.10.0. .. Becton Dickinson (BD) Reagent/resource Reference or source Identifier or catalog number FIJI is just ImageJ ImageJ, NIH GraphPad Prism version 10 GraphPad Gene set enrichment analysis UC San Diego/Broad Institute https:// www.gseamsigdb.org/gsea/ index.jsp Other Cytek Aurora 5-Laser Spectral Cytometer Cytek Biosciences Nikon AX R with NSPARC confocal system Nikon
Activation Assay:Article Title: ER stress induced mitochondrial dysfunction drives Treg instability in coronary artery disease.
Article Snippet: Reagents and tools table Reagent/resource Reference or source Identifier or catalog number Experimental models Healthy human PBMC’s Clinical core, La Jolla Institute for Immunology (LJI), La Jolla, California, USA PBMCs from CAD patients University of Virginia Health System, Charlottesville, Virginia, USA Coronary Assessment in Virginia (CAVA) cohort Recombinant DNA Not applicable Antibodies Alexa Fluor® 700 antihuman CD8a, Clone RPAT8 Biolegend Cat.#301028, 1:100 Pacific BlueTM anti-human CD4, Clone RPA-T4 Biolegend Cat. #300524, 1:100 10 EMBO Molecular Medicine © The Author(s) D ow nloaded from https://w w w .em bopress.org on N ovem ber 8, 2025 from IP 182.10.130.72. .. Reagent/resource Reference or source Identifier or catalog number Bovine serum albumin Cell Signaling Cat. #9998S Blotting Grade Blocker Non-Fat Dry Milk Biorad Cat. #1706404XTU TGXTM FastCastTM Acrylamide Kit, 12% Biorad Cat. #1610175 Ammonium persulfate Sigma Cat. #A3678 TEMED Biorad Cat. #1610800 RIPA Lysis buffer Cell Signaling Cat. #9806 Protease Inhibitor Roche Cat.#11697498001 4X Laemmli sample buffer Biorad Cat.#1610747 2-Mercaptoethanol ThermoFisher Cat.#21985023 SuperSignalTM West Femto Maximum Sensitivity Substrate 100ml ThermoFisher Cat.#34095 DynabeadsTM Human T-Activator CD3/CD28 for T Cell Expansion and Activation ThermoFisher Cat.#11161D TexMACS Miltenyi Biotec Cat.#130-097-196 RPMI 1640 Medium, no phenol red ThermoFisher Cat.#11835030 Puromycin Sigma Cat.#P7255 2-Deoxy-D-Glucose Sigma Cat.#D6134 Oligomycin A Sigma Cat.#75351 eBioscienceTM Fixation/ Permeabilization Concentrate ThermoFisher Cat.#00-5123-43 eBioscienceTM Fixation/ Permeabilization Diluent ThermoFisher Cat.#00-5223-56 eBioscienceTM Permeabilization Buffer (10X) ThermoFisher Cat.#00-8333-56 Glass-bottom dishes Cellvis Cat.#D35-20-1.5-N Poly D lysine ThermoFisher Cat.#A3890401 EasySepTM Human CD4+ T Cell Isolation Kit STEMCELL Technologies Cat.#17952 EasySepTM Human Naïve CD4+ T Cell Isolation Kit STEMCELL Technologies Cat.#19555 EasySepTM Human CD4+CD127lowCD25+ Regulatory T Cell Isolation Kit STEMCELL Technologies Cat.#18063 ImmunoCultTM-XF T Cell Expansion Medium STEMCELL Technologies Cat.#10981 ImmunoCultTM Human Treg Differentiation Supplement STEMCELL Technologies Cat.#10977 1×Cell Stimulation Cocktail, eBioscience ThermoFisher Cat.# 00-4970-93 1× Protein Transport Inhibitor Cocktail, eBioscience ThermoFisher Cat.# 00-4980-93 Software FlowJo version 10.10.0. .. Becton Dickinson (BD) Reagent/resource Reference or source Identifier or catalog number FIJI is just ImageJ ImageJ, NIH GraphPad Prism version 10 GraphPad Gene set enrichment analysis UC San Diego/Broad Institute https:// www.gseamsigdb.org/gsea/ index.jsp Other Cytek Aurora 5-Laser Spectral Cytometer Cytek Biosciences Nikon AX R with NSPARC confocal system Nikon
Cell Isolation:Article Title: ER stress induced mitochondrial dysfunction drives Treg instability in coronary artery disease.
Article Snippet: Reagents and tools table Reagent/resource Reference or source Identifier or catalog number Experimental models Healthy human PBMC’s Clinical core, La Jolla Institute for Immunology (LJI), La Jolla, California, USA PBMCs from CAD patients University of Virginia Health System, Charlottesville, Virginia, USA Coronary Assessment in Virginia (CAVA) cohort Recombinant DNA Not applicable Antibodies Alexa Fluor® 700 antihuman CD8a, Clone RPAT8 Biolegend Cat.#301028, 1:100 Pacific BlueTM anti-human CD4, Clone RPA-T4 Biolegend Cat. #300524, 1:100 10 EMBO Molecular Medicine © The Author(s) D ow nloaded from https://w w w .em bopress.org on N ovem ber 8, 2025 from IP 182.10.130.72. .. Reagent/resource Reference or source Identifier or catalog number Bovine serum albumin Cell Signaling Cat. #9998S Blotting Grade Blocker Non-Fat Dry Milk Biorad Cat. #1706404XTU TGXTM FastCastTM Acrylamide Kit, 12% Biorad Cat. #1610175 Ammonium persulfate Sigma Cat. #A3678 TEMED Biorad Cat. #1610800 RIPA Lysis buffer Cell Signaling Cat. #9806 Protease Inhibitor Roche Cat.#11697498001 4X Laemmli sample buffer Biorad Cat.#1610747 2-Mercaptoethanol ThermoFisher Cat.#21985023 SuperSignalTM West Femto Maximum Sensitivity Substrate 100ml ThermoFisher Cat.#34095 DynabeadsTM Human T-Activator CD3/CD28 for T Cell Expansion and Activation ThermoFisher Cat.#11161D TexMACS Miltenyi Biotec Cat.#130-097-196 RPMI 1640 Medium, no phenol red ThermoFisher Cat.#11835030 Puromycin Sigma Cat.#P7255 2-Deoxy-D-Glucose Sigma Cat.#D6134 Oligomycin A Sigma Cat.#75351 eBioscienceTM Fixation/ Permeabilization Concentrate ThermoFisher Cat.#00-5123-43 eBioscienceTM Fixation/ Permeabilization Diluent ThermoFisher Cat.#00-5223-56 eBioscienceTM Permeabilization Buffer (10X) ThermoFisher Cat.#00-8333-56 Glass-bottom dishes Cellvis Cat.#D35-20-1.5-N Poly D lysine ThermoFisher Cat.#A3890401 EasySepTM Human CD4+ T Cell Isolation Kit STEMCELL Technologies Cat.#17952 EasySepTM Human Naïve CD4+ T Cell Isolation Kit STEMCELL Technologies Cat.#19555 EasySepTM Human CD4+CD127lowCD25+ Regulatory T Cell Isolation Kit STEMCELL Technologies Cat.#18063 ImmunoCultTM-XF T Cell Expansion Medium STEMCELL Technologies Cat.#10981 ImmunoCultTM Human Treg Differentiation Supplement STEMCELL Technologies Cat.#10977 1×Cell Stimulation Cocktail, eBioscience ThermoFisher Cat.# 00-4970-93 1× Protein Transport Inhibitor Cocktail, eBioscience ThermoFisher Cat.# 00-4980-93 Software FlowJo version 10.10.0. .. Becton Dickinson (BD) Reagent/resource Reference or source Identifier or catalog number FIJI is just ImageJ ImageJ, NIH GraphPad Prism version 10 GraphPad Gene set enrichment analysis UC San Diego/Broad Institute https:// www.gseamsigdb.org/gsea/ index.jsp Other Cytek Aurora 5-Laser Spectral Cytometer Cytek Biosciences Nikon AX R with NSPARC confocal system Nikon
Software:Article Title: ER stress induced mitochondrial dysfunction drives Treg instability in coronary artery disease.
Article Snippet: Reagents and tools table Reagent/resource Reference or source Identifier or catalog number Experimental models Healthy human PBMC’s Clinical core, La Jolla Institute for Immunology (LJI), La Jolla, California, USA PBMCs from CAD patients University of Virginia Health System, Charlottesville, Virginia, USA Coronary Assessment in Virginia (CAVA) cohort Recombinant DNA Not applicable Antibodies Alexa Fluor® 700 antihuman CD8a, Clone RPAT8 Biolegend Cat.#301028, 1:100 Pacific BlueTM anti-human CD4, Clone RPA-T4 Biolegend Cat. #300524, 1:100 10 EMBO Molecular Medicine © The Author(s) D ow nloaded from https://w w w .em bopress.org on N ovem ber 8, 2025 from IP 182.10.130.72. .. Reagent/resource Reference or source Identifier or catalog number Bovine serum albumin Cell Signaling Cat. #9998S Blotting Grade Blocker Non-Fat Dry Milk Biorad Cat. #1706404XTU TGXTM FastCastTM Acrylamide Kit, 12% Biorad Cat. #1610175 Ammonium persulfate Sigma Cat. #A3678 TEMED Biorad Cat. #1610800 RIPA Lysis buffer Cell Signaling Cat. #9806 Protease Inhibitor Roche Cat.#11697498001 4X Laemmli sample buffer Biorad Cat.#1610747 2-Mercaptoethanol ThermoFisher Cat.#21985023 SuperSignalTM West Femto Maximum Sensitivity Substrate 100ml ThermoFisher Cat.#34095 DynabeadsTM Human T-Activator CD3/CD28 for T Cell Expansion and Activation ThermoFisher Cat.#11161D TexMACS Miltenyi Biotec Cat.#130-097-196 RPMI 1640 Medium, no phenol red ThermoFisher Cat.#11835030 Puromycin Sigma Cat.#P7255 2-Deoxy-D-Glucose Sigma Cat.#D6134 Oligomycin A Sigma Cat.#75351 eBioscienceTM Fixation/ Permeabilization Concentrate ThermoFisher Cat.#00-5123-43 eBioscienceTM Fixation/ Permeabilization Diluent ThermoFisher Cat.#00-5223-56 eBioscienceTM Permeabilization Buffer (10X) ThermoFisher Cat.#00-8333-56 Glass-bottom dishes Cellvis Cat.#D35-20-1.5-N Poly D lysine ThermoFisher Cat.#A3890401 EasySepTM Human CD4+ T Cell Isolation Kit STEMCELL Technologies Cat.#17952 EasySepTM Human Naïve CD4+ T Cell Isolation Kit STEMCELL Technologies Cat.#19555 EasySepTM Human CD4+CD127lowCD25+ Regulatory T Cell Isolation Kit STEMCELL Technologies Cat.#18063 ImmunoCultTM-XF T Cell Expansion Medium STEMCELL Technologies Cat.#10981 ImmunoCultTM Human Treg Differentiation Supplement STEMCELL Technologies Cat.#10977 1×Cell Stimulation Cocktail, eBioscience ThermoFisher Cat.# 00-4970-93 1× Protein Transport Inhibitor Cocktail, eBioscience ThermoFisher Cat.# 00-4980-93 Software FlowJo version 10.10.0. .. Becton Dickinson (BD) Reagent/resource Reference or source Identifier or catalog number FIJI is just ImageJ ImageJ, NIH GraphPad Prism version 10 GraphPad Gene set enrichment analysis UC San Diego/Broad Institute https:// www.gseamsigdb.org/gsea/ index.jsp Other Cytek Aurora 5-Laser Spectral Cytometer Cytek Biosciences Nikon AX R with NSPARC confocal system Nikon
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