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Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV140106	1/2	Prochlorococcus	Bacteria	(d)(U)C DG Filtration UF	Biller SJ	2014	57%
Study summary Full title Bacterial vesicles in marine ecosystems All authors Biller SJ, Schubotz F, Roggensack SE, Thompson AW, Summons RE, Chisholm SW Journal Science Abstract Many heterotrophic bacteria are known to release extracellular vesicles, facilitating interactions b (show more...)Many heterotrophic bacteria are known to release extracellular vesicles, facilitating interactions between cells and their environment from a distance. Vesicle production has not been described in photoautotrophs, however, and the prevalence and characteristics of vesicles in natural ecosystems is unknown. Here, we report that cultures of Prochlorococcus, a numerically dominant marine cyanobacterium, continuously release lipid vesicles containing proteins, DNA, and RNA. We also show that vesicles carrying DNA from diverse bacteria are abundant in coastal and open-ocean seawater samples. Prochlorococcus vesicles can support the growth of heterotrophic bacterial cultures, which implicates these structures in marine carbon flux. The ability of vesicles to deliver diverse compounds in discrete packages adds another layer of complexity to the flow of information, energy, and biomolecules in marine microbial communities. (hide) EV-METRIC 57% (95th 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 Bacteria Sample origin NAY Focus vesicles Bacterial vesicles 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 (d)(U)C DG Filtration UF Adj. k-factor 256 (pelleting) Protein markers EV: non-EV: Proteomics yes Show all info Study aim Function Sample Species Prochlorococcus Sample Type Bacteria Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 100,000 g and 150,000 g Pelleting performed Yes Pelleting: time(min) 120 Pelleting: rotor type SW32 Pelleting: adjusted k-factor 256.0 Density gradient Only used for validation of main results Yes Highest density fraction 40 Orientation Bottom-up Rotor type SW60 Speed (g) 100000 Filtration steps 0.22µm or 0.2µm Characterization: Protein analysis Characterization: Particle analysis NTA EM EM-type transmission EM/ cryo EM/ scanning EM Image type Wide-field

	EV140106	2/2	Prochlorococcus	Coastal water	(d)(U)C DG Filtration UF	Biller SJ	2014	57%
Study summary Full title Bacterial vesicles in marine ecosystems All authors Biller SJ, Schubotz F, Roggensack SE, Thompson AW, Summons RE, Chisholm SW Journal Science Abstract Many heterotrophic bacteria are known to release extracellular vesicles, facilitating interactions b (show more...)Many heterotrophic bacteria are known to release extracellular vesicles, facilitating interactions between cells and their environment from a distance. Vesicle production has not been described in photoautotrophs, however, and the prevalence and characteristics of vesicles in natural ecosystems is unknown. Here, we report that cultures of Prochlorococcus, a numerically dominant marine cyanobacterium, continuously release lipid vesicles containing proteins, DNA, and RNA. We also show that vesicles carrying DNA from diverse bacteria are abundant in coastal and open-ocean seawater samples. Prochlorococcus vesicles can support the growth of heterotrophic bacterial cultures, which implicates these structures in marine carbon flux. The ability of vesicles to deliver diverse compounds in discrete packages adds another layer of complexity to the flow of information, energy, and biomolecules in marine microbial communities. (hide) EV-METRIC 57% (50th 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 Coastal water Sample origin NAY Focus vesicles Bacterial vesicles 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 (d)(U)C DG Filtration UF Adj. k-factor 256 (pelleting) Protein markers EV: non-EV: Proteomics yes Show all info Study aim Function Sample Species Prochlorococcus Sample Type Coastal water Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 100,000 g and 150,000 g Pelleting performed Yes Pelleting: time(min) 120 Pelleting: rotor type SW32 Pelleting: adjusted k-factor 256.0 Density gradient Only used for validation of main results Yes Highest density fraction 40 Orientation Bottom-up Rotor type SW60 Speed (g) 100000 Filtration steps 0.22µm or 0.2µm Characterization: Protein analysis Characterization: Particle analysis NTA EM EM-type transmission EM/ cryo EM/ scanning EM Image type Wide-field

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EV-TRACK ID	EV140106
species	Prochlorococcus
sample type	Bacteria	Coastal water
condition	NAY	NAY
separation protocol	(d)(U)C DG Filtration UF	(d)(U)C DG Filtration UF
Exp. nr.	1	2
EV-METRIC %	57	57