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Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV200192	1/2	Exophiala dermatitidis	EXF-10123	(d)(U)C Filtration UF DG	Lavrin, Teja	2020	58%
Study summary Full title The Neurotropic Black Yeast Exophiala dermatitidis Induces Neurocytotoxicity in Neuroblastoma Cells and Progressive Cell Death All authors Teja Lavrin, Tilen Konte, Rok Kostanjšek, Simona Sitar, Kristina Sepčič, Sonja Prpar Mihevc, Ema Žagar, Vera Župunski, Metka Lenassi, Boris Rogelj, Nina Gunde Cimerman Journal Cells Abstract The neurotropic and extremophilic black yeast Exophiala dermatitidis (Herpotrichellaceae) inhabits d (show more...)The neurotropic and extremophilic black yeast Exophiala dermatitidis (Herpotrichellaceae) inhabits diverse indoor environments, in particular bathrooms, steam baths, and dishwashers. Here, we show that the selected strain, EXF-10123, is polymorphic, can grow at 37 °C, is able to assimilate aromatic hydrocarbons (toluene, mineral oil, n-hexadecane), and shows abundant growth with selected neurotransmitters (acetylcholine, gamma-aminobutyric acid, glycine, glutamate, and dopamine) as sole carbon sources. We have for the first time demonstrated the effect of E. dermatitidis on neuroblastoma cell model SH-SY5Y. Aqueous and organic extracts of E. dermatitidis biomass reduced SH-SY5Y viability by 51% and 37%, respectively. Melanized extracellular vesicles (EVs) prepared from this strain reduced viability of the SH-SY5Y to 21%, while non-melanized EVs were considerably less neurotoxic (79% viability). We also demonstrated direct interactions of E. dermatitidis with SH-SY5Y by scanning electron and confocal fluorescence microscopy. The observed invasion and penetration of neuroblastoma cells by E. dermatitidis hyphae presumably causes the degradation of most neuroblastoma cells in only three days. This may represent a so far unknown indirect or direct cause for the development of some neurodegenerative diseases such as Alzheimer's. (hide) EV-METRIC 58% (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 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 Filtration Ultrafiltration Density gradient Protein markers EV: None non-EV: None Proteomics no EV density (g/ml) 1.09–1.32 Show all info Study aim Function Sample Species Exophiala dermatitidis Sample Type Cell culture supernatant EV-producing cells EXF-10123 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 Pelleting performed Yes Pelleting: time(min) 70 Pelleting: rotor type MLA-50 Pelleting: speed (g) 100000 Wash: volume per pellet (ml) Not specified Wash: time (min) 70 Wash: Rotor Type TLA-55 Wash: speed (g) 100000 Density gradient Only used for validation of main results Yes Type Continuous Lowest density fraction 20 Highest density fraction 60 Total gradient volume, incl. sample (mL) 4.8 Sample volume (mL) 0.4 Orientation Not specified Rotor type Not specified Speed (g) 100000 Duration (min) 1080 Fraction volume (mL) 0.4 Fraction processing Precipitation Filtration steps 0.22µm or 0.2µm Ultra filtration Cut-off size (kDa) 100 Membrane type Not specified Other Name other separation method Filtration Other Name other separation method Ultrafiltration Other Name other separation method Density gradient Characterization: Protein analysis None Protein Concentration Method BCA Characterization: Lipid analysis No Characterization: Particle analysis Particle analysis: flow cytometry Hardware adjustment EM EM-type Transmission-EM Image type Close-up Report type Mean Report size 90 EV-concentration Yes

	EV200192	2/2	Exophiala dermatitidis	EXF-10123	(d)(U)C Filtration UF DG	Lavrin, Teja	2020	58%
Study summary Full title The Neurotropic Black Yeast Exophiala dermatitidis Induces Neurocytotoxicity in Neuroblastoma Cells and Progressive Cell Death All authors Teja Lavrin, Tilen Konte, Rok Kostanjšek, Simona Sitar, Kristina Sepčič, Sonja Prpar Mihevc, Ema Žagar, Vera Župunski, Metka Lenassi, Boris Rogelj, Nina Gunde Cimerman Journal Cells Abstract The neurotropic and extremophilic black yeast Exophiala dermatitidis (Herpotrichellaceae) inhabits d (show more...)The neurotropic and extremophilic black yeast Exophiala dermatitidis (Herpotrichellaceae) inhabits diverse indoor environments, in particular bathrooms, steam baths, and dishwashers. Here, we show that the selected strain, EXF-10123, is polymorphic, can grow at 37 °C, is able to assimilate aromatic hydrocarbons (toluene, mineral oil, n-hexadecane), and shows abundant growth with selected neurotransmitters (acetylcholine, gamma-aminobutyric acid, glycine, glutamate, and dopamine) as sole carbon sources. We have for the first time demonstrated the effect of E. dermatitidis on neuroblastoma cell model SH-SY5Y. Aqueous and organic extracts of E. dermatitidis biomass reduced SH-SY5Y viability by 51% and 37%, respectively. Melanized extracellular vesicles (EVs) prepared from this strain reduced viability of the SH-SY5Y to 21%, while non-melanized EVs were considerably less neurotoxic (79% viability). We also demonstrated direct interactions of E. dermatitidis with SH-SY5Y by scanning electron and confocal fluorescence microscopy. The observed invasion and penetration of neuroblastoma cells by E. dermatitidis hyphae presumably causes the degradation of most neuroblastoma cells in only three days. This may represent a so far unknown indirect or direct cause for the development of some neurodegenerative diseases such as Alzheimer's. (hide) EV-METRIC 58% (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 Cell culture supernatant Sample origin Melanin inhibitor 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 Filtration Ultrafiltration Density gradient Protein markers EV: None non-EV: None Proteomics no EV density (g/ml) 1.09–1.32 Show all info Study aim Function Sample Species Exophiala dermatitidis Sample Type Cell culture supernatant EV-producing cells EXF-10123 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 Pelleting performed Yes Pelleting: time(min) 70 Pelleting: rotor type MLA-50 Pelleting: speed (g) 100000 Wash: volume per pellet (ml) Not specified Wash: time (min) 70 Wash: Rotor Type TLA-55 Wash: speed (g) 100000 Density gradient Only used for validation of main results Yes Type Continuous Lowest density fraction 20 Highest density fraction 60 Total gradient volume, incl. sample (mL) 4.8 Sample volume (mL) 0.4 Orientation Not specified Rotor type Not specified Speed (g) 100000 Duration (min) 1080 Fraction volume (mL) 0.4 Fraction processing Precipitation Filtration steps 0.22µm or 0.2µm Ultra filtration Cut-off size (kDa) 100 Membrane type Not specified Other Name other separation method Filtration Other Name other separation method Ultrafiltration Other Name other separation method Density gradient Characterization: Protein analysis None Protein Concentration Method BCA Characterization: Lipid analysis No Characterization: Particle analysis Particle analysis: flow cytometry Hardware adjustment EM EM-type Transmission-EM Image type Close-up Report type Mean Report size 75 EV-concentration Yes

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EV-TRACK ID	EV200192
species	Exophiala dermatitidis
sample type	Cell culture
cell type	EXF-10123
condition	Control condition	Melanin inhibitor
separation protocol	dUC Filtration Ultrafiltration Density gradient	dUC Filtration Ultrafiltration Density gradient
Exp. nr.	1	2
EV-METRIC %	58	58