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Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV230967	1/1	Eimeria falciformis	NA	(d)(U)C DG UF Filtration	Olajide JS	2022	67%
Study summary Full title Eimeria falciformis secretes extracellular vesicles to modulate proinflammatory response during interaction with mouse intestinal epithelial cells. All authors Olajide JS, Xiong L, Yang S, Qu Z, Xu X, Yang B, Wang J, Liu B, Ma X, Cai J Journal Parasit Vectors Abstract Protozoan parasite secretions can be triggered by various modified media and diverse physicochemical (show more...)Protozoan parasite secretions can be triggered by various modified media and diverse physicochemical stressors. Equally, host-parasite interactions are known to co-opt the exchange and secretion of soluble biochemical components. Analysis of Eimeria falciformis sporozoite secretions in response to interaction with mouse intestinal epithelial cells (MIECs) may reveal parasite secretory motifs, protein composition and inflammatory activities of E. falciformis extracellular vesicles (EVs). (hide) EV-METRIC 67% (94th 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 extracellular 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 Ultrafiltration Filtration Adj. k-factor 58515 (pelleting) / 58515 (washing) Protein markers EV: HSP70/ HSP90 non-EV: None Proteomics yes EV density (g/ml) 1.12-1.179 Show all info Study aim Biogenesis/cargo sorting/Mechanism of uptake/transfer/Identification of content (omics approaches) Sample Species Eimeria falciformis Sample Type Cell culture supernatant EV-harvesting Medium EV-depleted medium Preparation of EDS Commercial EDS Cell viability (%) 100 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: rotor type T-890 Pelleting: speed (g) 120,000 Pelleting: adjusted k-factor 58515 Wash: volume per pellet (ml) 1 Wash: time (min) 240 Wash: Rotor Type T-890 Wash: speed (g) 120000 Wash: adjusted k-factor 58515 Density gradient Only used for validation of main results Yes Type Discontinuous Number of initial discontinuous layers 5 Lowest density fraction 5% Highest density fraction 50% Total gradient volume, incl. sample (mL) 7.4 Sample volume (mL) 1 Orientation Top-down Speed (g) 120,000 Duration (min) 1,200 Fraction volume (mL) 0.2 Fraction processing Centrifugation Pelleting: volume per fraction 0.2 Pelleting: speed (g) 120,000 Pelleting: adjusted k-factor 58515 Pelleting-wash: volume per pellet (mL) 0.1 Pelleting-wash: duration (min) 240 Pelleting-wash: speed (g) T-890 Filtration steps 0.2 or 0.22 ?m Ultra filtration Cut-off size (kDa) 10 Membrane type Polyethersulfone (PES) Characterization: Protein analysis Protein Concentration Method BCA Western Blot Detected EV-associated proteins HSP70 Not detected EV-associated proteins HSP90 Characterization: RNA analysis RNA analysis Type Capillary electrophoresis (e.g. Bioanalyzer) Proteinase treatment No RNAse treatment No Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Mean Reported size (nm) 246 ± 2 EV concentration Yes EM EM-type Transmission-EM Image type Wide-field

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EV-TRACK ID	EV230967
species	Eimeria falciformis
sample type	Cell culture
condition	Control condition
separation protocol	dUC/ Density gradient/ Ultrafiltration/ Filtration
Exp. nr.	1
EV-METRIC %	67