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Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV230725	1/1	Escherichia coli	536 (O6:K15:H31)	(d)(U)C DG Filtration UF	Blenkiron C	2016	29%
Study summary Full title Uropathogenic Escherichia coli Releases Extracellular Vesicles That Are Associated with RNA. All authors Blenkiron C, Simonov D, Muthukaruppan A, Tsai P, Dauros P, Green S, Hong J, Print CG, Swift S, Phillips AR Journal PLoS One Abstract Bacterium-to-host signalling during infection is a complex process involving proteins, lipids and ot (show more...)Bacterium-to-host signalling during infection is a complex process involving proteins, lipids and other diffusible signals that manipulate host cell biology for pathogen survival. Bacteria also release membrane vesicles (MV) that can carry a cargo of effector molecules directly into host cells. Supported by recent publications, we hypothesised that these MVs also associate with RNA, which may be directly involved in the modulation of the host response to infection. (hide) EV-METRIC 29% (67th 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 Control condition Focus vesicles Membrane vesicle (MV) 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 Ultrafiltration Protein markers EV: None non-EV: None Proteomics no Show all info Study aim Function/Identification of content (omics approaches) Sample Species Escherichia coli Sample Type Cell culture supernatant EV-producing cells 536 (O6:K15:H31) 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 Between 50,000 g and 100,000 g Pelleting performed Yes Pelleting: speed (g) 75000 Density gradient Type Continuous Lowest density fraction 20% Highest density fraction 45% Total gradient volume, incl. sample (mL) 10.8 Speed (g) 100000 Duration (min) 960 Fraction volume (mL) Ultrafiltration Fraction processing None Filtration steps 0.22µm or 0.2µm Ultra filtration Cut-off size (kDa) 100 Membrane type NS Characterization: Protein analysis None Protein Concentration Method BCA Characterization: RNA analysis RNA analysis Type (RT)(q)PCR/ RNA sequencing Proteinase treatment No RNAse treatment Yes Characterization: Lipid analysis No Characterization: Particle analysis NTA EV concentration Yes Particle yield as number of particles per milliliter of starting sample: 2.00E+09 EM EM-type Transmission-EM Image type Wide-field

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EV-TRACK ID	EV230725
species	Escherichia coli
sample type	Cell culture

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