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Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV130090	1/2	Rattus norvegicus/rattus	NAY	(d)(U)C DG	Grapp M	2013	33%
Study summary Full title Choroid plexus transcytosis and exosome shuttling deliver folate into brain parenchyma All authors Grapp M, Wrede A, Schweizer M, Hüwel S, Galla HJ, Snaidero N, Simons M, Bückers J, Low PS, Urlaub H, Gärtner J, Steinfeld R Journal Nat Commun Abstract Loss of folate receptor-? function is associated with cerebral folate transport deficiency and child (show more...)Loss of folate receptor-? function is associated with cerebral folate transport deficiency and childhood-onset neurodegeneration. To clarify the mechanism of cerebral folate transport at the blood-cerebrospinal fluid barrier, we investigate the transport of 5-methyltetrahydrofolate in polarized cells. Here we identify folate receptor-?-positive intralumenal vesicles within multivesicular bodies and demonstrate the directional cotransport of human folate receptor-?, and labelled folate from the basolateral to the apical membrane in rat choroid plexus cells. Both the apical medium of folate receptor-?-transfected rat choroid plexus cells and human cerebrospinal fluid contain folate receptor-?-positive exosomes. Loss of folate receptor-?-expressing cerebrospinal fluid exosomes correlates with severely reduced 5-methyltetrahydrofolate concentration, corroborating the importance of the folate receptor-?-mediated folate transport in the cerebrospinal fluid. Intraventricular injections of folate receptor-?-positive and -negative exosomes into mouse brains demonstrate folate receptor-?-dependent delivery of exosomes into the brain parenchyma. Our results unravel a new pathway of folate receptor-?-dependent exosome-mediated folate delivery into the brain parenchyma and opens new avenues for cerebral drug targeting. (hide) EV-METRIC 33% (75th 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 NAY Focus vesicles exosomes 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 Protein markers EV: Alix/ Flotillin2 non-EV: Proteomics yes EV density (g/ml) 1.13-1.16 Show all info Study aim Function Sample Species Rattus norvegicus/rattus Sample Type Cell culture supernatant Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 800 g and 10,000 g Between 10,000 g and 50,000 g Between 100,000 g and 150,000 g Pelleting performed Yes Pelleting: time(min) 150 Density gradient Only used for validation of main results Yes Orientation Top-down Characterization: Protein analysis Western Blot Antibody details provided? No Detected EV-associated proteins Alix/ Flotillin2 ELISA Antibody details provided? No Detected EV-associated proteins Flotillin2

	EV130090	2/2	Homo sapiens	"Cerebrospinal fluid"	(d)(U)C DG	Grapp M	2013	33%
Study summary Full title Choroid plexus transcytosis and exosome shuttling deliver folate into brain parenchyma All authors Grapp M, Wrede A, Schweizer M, Hüwel S, Galla HJ, Snaidero N, Simons M, Bückers J, Low PS, Urlaub H, Gärtner J, Steinfeld R Journal Nat Commun Abstract Loss of folate receptor-? function is associated with cerebral folate transport deficiency and child (show more...)Loss of folate receptor-? function is associated with cerebral folate transport deficiency and childhood-onset neurodegeneration. To clarify the mechanism of cerebral folate transport at the blood-cerebrospinal fluid barrier, we investigate the transport of 5-methyltetrahydrofolate in polarized cells. Here we identify folate receptor-?-positive intralumenal vesicles within multivesicular bodies and demonstrate the directional cotransport of human folate receptor-?, and labelled folate from the basolateral to the apical membrane in rat choroid plexus cells. Both the apical medium of folate receptor-?-transfected rat choroid plexus cells and human cerebrospinal fluid contain folate receptor-?-positive exosomes. Loss of folate receptor-?-expressing cerebrospinal fluid exosomes correlates with severely reduced 5-methyltetrahydrofolate concentration, corroborating the importance of the folate receptor-?-mediated folate transport in the cerebrospinal fluid. Intraventricular injections of folate receptor-?-positive and -negative exosomes into mouse brains demonstrate folate receptor-?-dependent delivery of exosomes into the brain parenchyma. Our results unravel a new pathway of folate receptor-?-dependent exosome-mediated folate delivery into the brain parenchyma and opens new avenues for cerebral drug targeting. (hide) EV-METRIC 33% (87th 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 "Cerebrospinal fluid" Sample origin NAY Focus vesicles exosomes 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 Protein markers EV: Flotillin2 non-EV: Proteomics yes EV density (g/ml) 1.11-1.16 Show all info Study aim Function Sample Species Homo sapiens Sample Type "Cerebrospinal fluid" Separation Method (Differential) (ultra)centrifugation dUC: centrifugation steps Between 800 g and 10,000 g Between 10,000 g and 50,000 g Between 100,000 g and 150,000 g Pelleting performed Yes Pelleting: time(min) 150 Density gradient Only used for validation of main results Yes Orientation Top-down Characterization: Protein analysis Western Blot Antibody details provided? No Detected EV-associated proteins Flotillin2 ELISA Antibody details provided? No Detected EV-associated proteins Flotillin2 Characterization: Particle analysis EM EM-type immune EM EM protein Flotillin2 Image type Close-up

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EV-TRACK ID	EV130090
species	Rattus norvegicus/rattus	Homo sapiens
sample type	Cell culture	Cerebrospinal fluid
cell type	NAY	NA
condition	NAY	NAY
separation protocol	(d)(U)C DG	(d)(U)C DG
Exp. nr.	1	2
EV-METRIC %	33	33