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Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV200048	1/2	Gallus gallus	semen	IAF (d)(U)C ExoQuick Filtration	Sheng Chen	2019	50%
Study summary Full title MicroRNA expression profile in extracellular vesicles derived from ALV-J infected chicken semen All authors Sheng Chen, Liqin Liao, Qiqi Zhao, Xinheng Zhang, Hongxin Li, Wencheng Lin, Feng Chen, Qingmei Xie Journal Virus Res Abstract MicroRNAs(miRNAs) have been reported to regulate gene expression in many processes. MiRNA in extrace (show more...)MicroRNAs(miRNAs) have been reported to regulate gene expression in many processes. MiRNA in extracellular vesicles (EVs) also have been widely investigated, while there is no studies of miRNAs in seminal EVs. Subgroup J of Avian leukosis virus (ALV-J) can be transmitted vertically, but the mechanism of it is not clear enough. This study was to examine the miRNA expression profile in seminal EVs and inquire into the relation between it and the vertical transmission by performing gene ontology (GO) and pathway enrichment analysis. Here, we first isolated and characterized seminal EVs by Nanoparticle Tracking Analysis、Western Blot and Transmission electron microscopy experiments. By deep sequencing of each EVs miRNA library, 9 typical differentially expressed miRNA, including 6 up-regulated and 3 down-regulated, were identified. Gene target prediction, GO annotation and KEGG pathway enrichment analysis showed possible function associated with these miRNAs. Overall, these findings will increase our understanding of the content and composition of miRNA in seminal EVs and provide new insights into the important role of the seminal EVs miRNAs regulation in ALV-J transmission. (hide) EV-METRIC 50% (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 semen 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 Immunoaffinity capture (non-commercial) (d)(U)C Commercial method Filtration Protein markers EV: CD81/ HSP70/ TSG101/ CD63/ CD9 non-EV: GRP78 Proteomics no Show all info Study aim NA Sample Species Gallus gallus Sample Type semen Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 800 g and 10,000 g Pelleting performed No Filtration steps 0.22µm or 0.2µm Commercial kit ExoQuick Immunoaffinity capture Selected surface protein(s) CD63 Characterization: Protein analysis Protein Concentration Method Not determined Western Blot Antibody details provided? Yes Antibody dilution provided? Yes Lysis buffer provided? Yes Detected EV-associated proteins CD9/ CD63/ TSG101/ HSP70/ CD81 Not detected EV-associated proteins HSP70/ CD81/ TSG101/ CD63/ CD9 Not detected contaminants GRP78 ELISA Antibody details provided? No Detected EV-associated proteins HSP70/ CD63/ CD81/ CD9/ TSG101 Characterization: RNA analysis RNA analysis Type (RT)(q)PCR;RNAsequencing Database No Proteinase treatment No RNAse treatment No Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Size range/distribution Reported size (nm) 110 EV concentration Yes Particle yield particles/ml;Yes, other: 858000000

	EV200048	2/2	Gallus gallus	semen	IAF (d)(U)C ExoQuick Filtration	Sheng Chen	2019	38%
Study summary Full title MicroRNA expression profile in extracellular vesicles derived from ALV-J infected chicken semen All authors Sheng Chen, Liqin Liao, Qiqi Zhao, Xinheng Zhang, Hongxin Li, Wencheng Lin, Feng Chen, Qingmei Xie Journal Virus Res Abstract MicroRNAs(miRNAs) have been reported to regulate gene expression in many processes. MiRNA in extrace (show more...)MicroRNAs(miRNAs) have been reported to regulate gene expression in many processes. MiRNA in extracellular vesicles (EVs) also have been widely investigated, while there is no studies of miRNAs in seminal EVs. Subgroup J of Avian leukosis virus (ALV-J) can be transmitted vertically, but the mechanism of it is not clear enough. This study was to examine the miRNA expression profile in seminal EVs and inquire into the relation between it and the vertical transmission by performing gene ontology (GO) and pathway enrichment analysis. Here, we first isolated and characterized seminal EVs by Nanoparticle Tracking Analysis、Western Blot and Transmission electron microscopy experiments. By deep sequencing of each EVs miRNA library, 9 typical differentially expressed miRNA, including 6 up-regulated and 3 down-regulated, were identified. Gene target prediction, GO annotation and KEGG pathway enrichment analysis showed possible function associated with these miRNAs. Overall, these findings will increase our understanding of the content and composition of miRNA in seminal EVs and provide new insights into the important role of the seminal EVs miRNAs regulation in ALV-J transmission. (hide) EV-METRIC 38% (68th 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 semen Sample origin ALV-J infected 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 Immunoaffinity capture (non-commercial) (d)(U)C Commercial method Filtration Protein markers EV: TSG101/ CD63/ CD81/ GRP78/ HSP70/ CD9 non-EV: None Proteomics no Show all info Study aim NA Sample Species Gallus gallus Sample Type semen Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 800 g and 10,000 g Pelleting performed No Filtration steps 0.22µm or 0.2µm Commercial kit ExoQuick Immunoaffinity capture Selected surface protein(s) CD63 Characterization: Protein analysis Protein Concentration Method Not determined Western Blot Antibody details provided? Yes Antibody dilution provided? Yes Lysis buffer provided? Yes Detected EV-associated proteins CD9/ CD63/ TSG101/ HSP70/ CD81 Not detected EV-associated proteins HSP70/ CD81/ GRP78/ TSG101/ CD63/ CD9 ELISA Antibody details provided? No Detected EV-associated proteins HSP70/ CD63/ CD81/ CD9/ TSG101 Characterization: RNA analysis RNA analysis Type (RT)(q)PCR;RNAsequencing Database No Proteinase treatment No RNAse treatment No Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Size range/distribution Reported size (nm) 127 EV concentration Yes Particle yield particles/ml;Yes, other: 8580000000

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EV-TRACK ID	EV200048
species	Gallus gallus
sample type	semen
condition	Control condition	ALV-J infected
separation protocol	IAF capture (non-commercial) (d)(U)C ExoQuick Filtration	IAF capture (non-commercial) (d)(U)C ExoQuick Filtration
Exp. nr.	1	2
EV-METRIC %	50	38