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Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV230850	1/6	Staphylococcus aureus	MSSA476	(d)(U)C DG Filtration	Askarian F	2018	57%
Study summary Full title Membrane-Derived Vesicles Promote Bacterial Virulence and Confer Protective Immunity in Murine Infection Models. All authors Askarian F, Lapek JD, Dongre M, Tsai CM, Kumaraswamy M, Kousha A, Valderrama JA, Ludviksen JA, Cavanagh JP, Uchiyama S, Mollnes TE, Gonzalez DJ, Wai SN, Nizet V, Johannessen M Journal Front Microbiol Abstract produces membrane-derived vesicles (MVs), which share functional properties to outer membrane vesicl (show more...)produces membrane-derived vesicles (MVs), which share functional properties to outer membrane vesicles. Atomic force microscopy revealed that -derived MVs are associated with the bacterial surface or released into the surrounding environment depending on bacterial growth conditions. By using a comparative proteomic approach, a total of 131 and 617 proteins were identified in MVs isolated from grown in Luria-Bertani and brain-heart infusion broth, respectively. Purified MVs derived from the bacteria grown in either media induced comparable levels of cytotoxicity and neutrophil-activation. Administration of exogenous MVs increased the resistance of to killing by whole blood or purified human neutrophils and increased survival . Finally, immunization of mice with -derived MVs induced production of IgM, total IgG, IgG1, IgG2a, and IgG2b resulting in protection against subcutaneous and systemic infection. Collectively, our results suggest MVs can influence bacterial-host interactions during systemic infections and provide protective immunity in murine models of infection. (hide) EV-METRIC 57% (92nd percentile of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type Cell culture supernatant Sample origin Luria Bertani broth Focus vesicles MV (membrane-derived vesicle) Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture (Differential) (ultra)centrifugation Density gradient Filtration Protein markers EV: None non-EV: None Proteomics yes Show all info Study aim Function Sample Species Staphylococcus aureus Sample Type Cell culture supernatant EV-producing cells MSSA476 EV-harvesting Medium Serum free medium Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 800 g and 10,000 g Between 100,000 g and 150,000 g Pelleting performed Yes Pelleting: time(min) 180-240 Pelleting: rotor type Type 45 Ti Pelleting: speed (g) 100000 Wash: time (min) 180-240 Wash: speed (g) 100000 Density gradient Only used for validation of main results Yes Type Discontinuous Number of initial discontinuous layers 7 Lowest density fraction 10% Highest density fraction 45% Total gradient volume, incl. sample (mL) 4,1 mL Sample volume (mL) 0.4 mL Orientation Top-down Speed (g) 180000 Duration (min) 180 Fraction volume (mL) 0.2 Fraction processing None Filtration steps 0.22µm or 0.2µm Characterization: Protein analysis Protein Concentration Method BCA Proteomics database No Characterization: Lipid analysis No Characterization: Particle analysis DLS Report type Mean Reported size (nm) 46.6 +- 5.9 EM EM-type Atomic force microscopy Image type Close-up

	EV230850	2/6	Staphylococcus aureus	MSSA476	(d)(U)C DG Filtration	Askarian F	2018	57%
Study summary Full title Membrane-Derived Vesicles Promote Bacterial Virulence and Confer Protective Immunity in Murine Infection Models. All authors Askarian F, Lapek JD, Dongre M, Tsai CM, Kumaraswamy M, Kousha A, Valderrama JA, Ludviksen JA, Cavanagh JP, Uchiyama S, Mollnes TE, Gonzalez DJ, Wai SN, Nizet V, Johannessen M Journal Front Microbiol Abstract produces membrane-derived vesicles (MVs), which share functional properties to outer membrane vesicl (show more...)produces membrane-derived vesicles (MVs), which share functional properties to outer membrane vesicles. Atomic force microscopy revealed that -derived MVs are associated with the bacterial surface or released into the surrounding environment depending on bacterial growth conditions. By using a comparative proteomic approach, a total of 131 and 617 proteins were identified in MVs isolated from grown in Luria-Bertani and brain-heart infusion broth, respectively. Purified MVs derived from the bacteria grown in either media induced comparable levels of cytotoxicity and neutrophil-activation. Administration of exogenous MVs increased the resistance of to killing by whole blood or purified human neutrophils and increased survival . Finally, immunization of mice with -derived MVs induced production of IgM, total IgG, IgG1, IgG2a, and IgG2b resulting in protection against subcutaneous and systemic infection. Collectively, our results suggest MVs can influence bacterial-host interactions during systemic infections and provide protective immunity in murine models of infection. (hide) EV-METRIC 57% (92nd percentile of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type Cell culture supernatant Sample origin Brain-heart infusion broth Focus vesicles MV (membrane-derived vesicle) Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture (Differential) (ultra)centrifugation Density gradient Filtration Protein markers EV: None non-EV: None Proteomics yes Show all info Study aim Function Sample Species Staphylococcus aureus Sample Type Cell culture supernatant EV-producing cells MSSA476 EV-harvesting Medium Serum free medium Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 800 g and 10,000 g Between 100,000 g and 150,000 g Pelleting performed Yes Pelleting: time(min) 180-240 Pelleting: rotor type Type 45 Ti Pelleting: speed (g) 100000 Wash: time (min) 180-240 Wash: speed (g) 100000 Density gradient Only used for validation of main results Yes Type Discontinuous Number of initial discontinuous layers 7 Lowest density fraction 10% Highest density fraction 45% Total gradient volume, incl. sample (mL) 4,1 mL Sample volume (mL) 0.4 mL Orientation Top-down Speed (g) 180000 Duration (min) 180 Fraction volume (mL) 0.2 Fraction processing None Filtration steps 0.22µm or 0.2µm Characterization: Protein analysis Protein Concentration Method BCA Proteomics database No Characterization: Lipid analysis No Characterization: Particle analysis DLS Report type Mean Reported size (nm) 24.4 +- 2.8 EM EM-type Atomic force microscopy Image type Close-up

	EV230850	3/6	Staphylococcus aureus	USA300 MRSA	(d)(U)C Filtration	Askarian F	2018	17%
Study summary Full title Membrane-Derived Vesicles Promote Bacterial Virulence and Confer Protective Immunity in Murine Infection Models. All authors Askarian F, Lapek JD, Dongre M, Tsai CM, Kumaraswamy M, Kousha A, Valderrama JA, Ludviksen JA, Cavanagh JP, Uchiyama S, Mollnes TE, Gonzalez DJ, Wai SN, Nizet V, Johannessen M Journal Front Microbiol Abstract produces membrane-derived vesicles (MVs), which share functional properties to outer membrane vesicl (show more...)produces membrane-derived vesicles (MVs), which share functional properties to outer membrane vesicles. Atomic force microscopy revealed that -derived MVs are associated with the bacterial surface or released into the surrounding environment depending on bacterial growth conditions. By using a comparative proteomic approach, a total of 131 and 617 proteins were identified in MVs isolated from grown in Luria-Bertani and brain-heart infusion broth, respectively. Purified MVs derived from the bacteria grown in either media induced comparable levels of cytotoxicity and neutrophil-activation. Administration of exogenous MVs increased the resistance of to killing by whole blood or purified human neutrophils and increased survival . Finally, immunization of mice with -derived MVs induced production of IgM, total IgG, IgG1, IgG2a, and IgG2b resulting in protection against subcutaneous and systemic infection. Collectively, our results suggest MVs can influence bacterial-host interactions during systemic infections and provide protective immunity in murine models of infection. (hide) EV-METRIC 17% (54th percentile of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type Cell culture supernatant Sample origin Luria Bertani broth Focus vesicles MV (membrane-derived vesicle) Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture (Differential) (ultra)centrifugation Filtration Protein markers EV: None non-EV: None Proteomics no Show all info Study aim Function Sample Species Staphylococcus aureus Sample Type Cell culture supernatant EV-producing cells USA300 MRSA EV-harvesting Medium Serum free medium Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 800 g and 10,000 g Between 100,000 g and 150,000 g Pelleting performed Yes Pelleting: time(min) 180-240 Pelleting: rotor type Type 45 Ti Pelleting: speed (g) 100000 Wash: time (min) 180-240 Wash: speed (g) 100000 Filtration steps 0.22µm or 0.2µm Characterization: Protein analysis Protein Concentration Method BCA Characterization: Lipid analysis No Characterization: Particle analysis DLS Report type Mean Reported size (nm) 46.6 +- 5.9 EM EM-type Atomic force microscopy Image type Close-up

	EV230850	4/6	Staphylococcus aureus	USA300 MRSA	(d)(U)C Filtration	Askarian F	2018	17%
Study summary Full title Membrane-Derived Vesicles Promote Bacterial Virulence and Confer Protective Immunity in Murine Infection Models. All authors Askarian F, Lapek JD, Dongre M, Tsai CM, Kumaraswamy M, Kousha A, Valderrama JA, Ludviksen JA, Cavanagh JP, Uchiyama S, Mollnes TE, Gonzalez DJ, Wai SN, Nizet V, Johannessen M Journal Front Microbiol Abstract produces membrane-derived vesicles (MVs), which share functional properties to outer membrane vesicl (show more...)produces membrane-derived vesicles (MVs), which share functional properties to outer membrane vesicles. Atomic force microscopy revealed that -derived MVs are associated with the bacterial surface or released into the surrounding environment depending on bacterial growth conditions. By using a comparative proteomic approach, a total of 131 and 617 proteins were identified in MVs isolated from grown in Luria-Bertani and brain-heart infusion broth, respectively. Purified MVs derived from the bacteria grown in either media induced comparable levels of cytotoxicity and neutrophil-activation. Administration of exogenous MVs increased the resistance of to killing by whole blood or purified human neutrophils and increased survival . Finally, immunization of mice with -derived MVs induced production of IgM, total IgG, IgG1, IgG2a, and IgG2b resulting in protection against subcutaneous and systemic infection. Collectively, our results suggest MVs can influence bacterial-host interactions during systemic infections and provide protective immunity in murine models of infection. (hide) EV-METRIC 17% (54th percentile of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type Cell culture supernatant Sample origin Brain-heart infusion broth Focus vesicles MV (membrane-derived vesicle) Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture (Differential) (ultra)centrifugation Filtration Protein markers EV: None non-EV: None Proteomics no Show all info Study aim Function Sample Species Staphylococcus aureus Sample Type Cell culture supernatant EV-producing cells USA300 MRSA EV-harvesting Medium Serum free medium Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 800 g and 10,000 g Between 100,000 g and 150,000 g Pelleting performed Yes Pelleting: time(min) 180-240 Pelleting: rotor type Type 45 Ti Pelleting: speed (g) 100000 Wash: time (min) 180-240 Wash: speed (g) 100000 Filtration steps 0.22µm or 0.2µm Characterization: Protein analysis Protein Concentration Method BCA Characterization: Lipid analysis No Characterization: Particle analysis DLS Report type Mean Reported size (nm) 24.4 +- 2.8 EM EM-type Atomic force microscopy Image type Close-up

	EV230850	5/6	Staphylococcus aureus	USA300 MRSA delHla	(d)(U)C Filtration	Askarian F	2018	17%
Study summary Full title Membrane-Derived Vesicles Promote Bacterial Virulence and Confer Protective Immunity in Murine Infection Models. All authors Askarian F, Lapek JD, Dongre M, Tsai CM, Kumaraswamy M, Kousha A, Valderrama JA, Ludviksen JA, Cavanagh JP, Uchiyama S, Mollnes TE, Gonzalez DJ, Wai SN, Nizet V, Johannessen M Journal Front Microbiol Abstract produces membrane-derived vesicles (MVs), which share functional properties to outer membrane vesicl (show more...)produces membrane-derived vesicles (MVs), which share functional properties to outer membrane vesicles. Atomic force microscopy revealed that -derived MVs are associated with the bacterial surface or released into the surrounding environment depending on bacterial growth conditions. By using a comparative proteomic approach, a total of 131 and 617 proteins were identified in MVs isolated from grown in Luria-Bertani and brain-heart infusion broth, respectively. Purified MVs derived from the bacteria grown in either media induced comparable levels of cytotoxicity and neutrophil-activation. Administration of exogenous MVs increased the resistance of to killing by whole blood or purified human neutrophils and increased survival . Finally, immunization of mice with -derived MVs induced production of IgM, total IgG, IgG1, IgG2a, and IgG2b resulting in protection against subcutaneous and systemic infection. Collectively, our results suggest MVs can influence bacterial-host interactions during systemic infections and provide protective immunity in murine models of infection. (hide) EV-METRIC 17% (54th percentile of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type Cell culture supernatant Sample origin Luria Bertani broth Focus vesicles MV (membrane-derived vesicle) Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture (Differential) (ultra)centrifugation Filtration Protein markers EV: None non-EV: None Proteomics no Show all info Study aim Function Sample Species Staphylococcus aureus Sample Type Cell culture supernatant EV-producing cells USA300 MRSA delHla EV-harvesting Medium Serum free medium Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 800 g and 10,000 g Between 100,000 g and 150,000 g Pelleting performed Yes Pelleting: time(min) 180-240 Pelleting: rotor type Type 45 Ti Pelleting: speed (g) 100000 Wash: time (min) 180-240 Wash: speed (g) 100000 Filtration steps 0.22µm or 0.2µm Characterization: Protein analysis Protein Concentration Method BCA Characterization: Lipid analysis No Characterization: Particle analysis DLS Report type Mean Reported size (nm) 46.6 +- 5.9 EM EM-type Atomic force microscopy Image type Close-up

	EV230850	6/6	Staphylococcus aureus	USA300 MRSA delHla	(d)(U)C Filtration	Askarian F	2018	17%
Study summary Full title Membrane-Derived Vesicles Promote Bacterial Virulence and Confer Protective Immunity in Murine Infection Models. All authors Askarian F, Lapek JD, Dongre M, Tsai CM, Kumaraswamy M, Kousha A, Valderrama JA, Ludviksen JA, Cavanagh JP, Uchiyama S, Mollnes TE, Gonzalez DJ, Wai SN, Nizet V, Johannessen M Journal Front Microbiol Abstract produces membrane-derived vesicles (MVs), which share functional properties to outer membrane vesicl (show more...)produces membrane-derived vesicles (MVs), which share functional properties to outer membrane vesicles. Atomic force microscopy revealed that -derived MVs are associated with the bacterial surface or released into the surrounding environment depending on bacterial growth conditions. By using a comparative proteomic approach, a total of 131 and 617 proteins were identified in MVs isolated from grown in Luria-Bertani and brain-heart infusion broth, respectively. Purified MVs derived from the bacteria grown in either media induced comparable levels of cytotoxicity and neutrophil-activation. Administration of exogenous MVs increased the resistance of to killing by whole blood or purified human neutrophils and increased survival . Finally, immunization of mice with -derived MVs induced production of IgM, total IgG, IgG1, IgG2a, and IgG2b resulting in protection against subcutaneous and systemic infection. Collectively, our results suggest MVs can influence bacterial-host interactions during systemic infections and provide protective immunity in murine models of infection. (hide) EV-METRIC 17% (54th percentile of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type Cell culture supernatant Sample origin Brain-heart infusion broth Focus vesicles MV (membrane-derived vesicle) Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture (Differential) (ultra)centrifugation Filtration Protein markers EV: None non-EV: None Proteomics no Show all info Study aim Function Sample Species Staphylococcus aureus Sample Type Cell culture supernatant EV-producing cells USA300 MRSA delHla EV-harvesting Medium Serum free medium Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 800 g and 10,000 g Between 100,000 g and 150,000 g Pelleting performed Yes Pelleting: time(min) 180-240 Pelleting: rotor type Type 45 Ti Pelleting: speed (g) 100000 Wash: time (min) 180-240 Wash: speed (g) 100000 Filtration steps 0.22µm or 0.2µm Characterization: Protein analysis Protein Concentration Method BCA Characterization: Lipid analysis No Characterization: Particle analysis DLS Report type Mean Reported size (nm) 24.4 +- 2.8 EM EM-type Atomic force microscopy Image type Close-up

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EV-TRACK ID	EV230850
species	Staphylococcus aureus
sample type	Cell culture
cell type	MSSA476	MSSA476	USA300 MRSA	USA300 MRSA	USA300 MRSA delHla	USA300 MRSA delHla
condition	Luria Bertani broth	Brain-heart infusion broth	Luria Bertani broth	Brain-heart infusion broth	Luria Bertani broth	Brain-heart infusion broth
separation protocol	dUC/ Density gradient/ Filtration	dUC/ Density gradient/ Filtration	dUC/ Filtration	dUC/ Filtration	dUC/ Filtration	dUC/ Filtration
Exp. nr.	1	2	3	4	5	6
EV-METRIC %	57	57	17	17	17	17