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Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV140236	1/2	Homo sapiens	HBE	(d)(U)C ExoQuick Filtration	Xu, Yuan	2015	25%
Study summary Full title Exosomal miR-21 derived from arsenite-transformed human bronchial epithelial cells promotes cell proliferation associated with arsenite carcinogenesis All authors Yuan Xu, Fei Luo, Yi Liu, Le Shi, Xiaolin Lu, Wenchao Xu, Qizhan Liu Journal Arch Toxicol Abstract Intercellular communications within the cancer microenvironment coordinate the assembly of various c (show more...)Intercellular communications within the cancer microenvironment coordinate the assembly of various cell types. Exosomes are mediators of intercellular communication in immune signaling, tumor promotion, stress responses, and angiogenesis. The present research aimed to determine whether miRNAs secreted from human bronchial epithelial (HBE) cells transformed by 1.0 μM arsenite are transferred into normal HBE cells and are functionally active in the recipient cells. The results show that miR-21 is involved in exosome-mediated intercellular communication between neoplastic and normal HBE cells. Exosomes derived from transformed HBE cells stimulated proliferation of normal HBE cells, whereas exosomes from miR-21 depleted cells failed to stimulate proliferation. In normal HBE cells, the expression of phosphatase and tensin homolog, a target gene for miR-21, was increased by exosomal miR-21, indicating that exogenous miRNAs, via exosomal transport, function-like endogenous miRNAs. Concordantly, specific reduction of miR-21 content in exosome-producing transformed cells abolished the stimulation of proliferation by exosomes. Collectively, the data indicate that transformed HBE cells release exosomes containing miR-21, stimulating proliferation in neighboring normal HBE cells and supporting the concept that exosomal miRNAs are involved in cell-cell communication during carcinogenesis induced by environmental chemicals. (hide) EV-METRIC 25% (64th 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 Arsenite-treated Focus vesicles exosome 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 ExoQuick Filtration Protein markers EV: CD63/ Flotillin1 non-EV: None Proteomics no Show all info Study aim Function Sample Species Homo sapiens Sample Type Cell culture supernatant EV-producing cells HBE EV-harvesting Medium Serum free medium 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 Other Name other separation method ExoQuick Characterization: Protein analysis Protein Concentration Method BCA Western Blot Antibody details provided? Yes Antibody dilution provided? Yes Detected EV-associated proteins Flotillin1/ CD63 Characterization: RNA analysis RNA analysis Type (RT)(q)PCR Proteinase treatment No RNAse treatment No Characterization: Lipid analysis No

	EV140236	2/2	Homo sapiens	HBE	(d)(U)C ExoQuick Filtration	Xu, Yuan	2015	25%
Study summary Full title Exosomal miR-21 derived from arsenite-transformed human bronchial epithelial cells promotes cell proliferation associated with arsenite carcinogenesis All authors Yuan Xu, Fei Luo, Yi Liu, Le Shi, Xiaolin Lu, Wenchao Xu, Qizhan Liu Journal Arch Toxicol Abstract Intercellular communications within the cancer microenvironment coordinate the assembly of various c (show more...)Intercellular communications within the cancer microenvironment coordinate the assembly of various cell types. Exosomes are mediators of intercellular communication in immune signaling, tumor promotion, stress responses, and angiogenesis. The present research aimed to determine whether miRNAs secreted from human bronchial epithelial (HBE) cells transformed by 1.0 μM arsenite are transferred into normal HBE cells and are functionally active in the recipient cells. The results show that miR-21 is involved in exosome-mediated intercellular communication between neoplastic and normal HBE cells. Exosomes derived from transformed HBE cells stimulated proliferation of normal HBE cells, whereas exosomes from miR-21 depleted cells failed to stimulate proliferation. In normal HBE cells, the expression of phosphatase and tensin homolog, a target gene for miR-21, was increased by exosomal miR-21, indicating that exogenous miRNAs, via exosomal transport, function-like endogenous miRNAs. Concordantly, specific reduction of miR-21 content in exosome-producing transformed cells abolished the stimulation of proliferation by exosomes. Collectively, the data indicate that transformed HBE cells release exosomes containing miR-21, stimulating proliferation in neighboring normal HBE cells and supporting the concept that exosomal miRNAs are involved in cell-cell communication during carcinogenesis induced by environmental chemicals. (hide) EV-METRIC 25% (64th 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 exosome 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 ExoQuick Filtration Protein markers EV: CD63/ Flotillin1 non-EV: None Proteomics no Show all info Study aim Function Sample Species Homo sapiens Sample Type Cell culture supernatant EV-producing cells HBE EV-harvesting Medium Serum free medium 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 Other Name other separation method ExoQuick Characterization: Protein analysis Protein Concentration Method BCA Western Blot Antibody details provided? Yes Antibody dilution provided? Yes Detected EV-associated proteins Flotillin1/ CD63 Characterization: RNA analysis RNA analysis Type (RT)(q)PCR Proteinase treatment No RNAse treatment No Characterization: Lipid analysis No

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EV-TRACK ID	EV140236
species	Homo sapiens
sample type	Cell culture
cell type	HBE
condition	Arsenite-treated	Control condition
separation protocol	(d)(U)C ExoQuick Filtration	(d)(U)C ExoQuick Filtration
Exp. nr.	1	2
EV-METRIC %	25	25